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Volume 1 - Number 1 May - September 1997
Volume 15 - Number 1 January 2011
The PDF version of the Atlas of Genetics and Cytogenetics in Oncology and Haematology is a reissue of the original articles published in collaboration with
the Institute for Scientific and Technical Information (INstitut de l’Information Scientifique et Technique - INIST) of the French National Center for Scientific
Research (CNRS) on its electronic publishing platform I-Revues.
Online and PDF versions of the Atlas of Genetics and Cytogenetics in Oncology and Haematology are hosted by INIST-CNRS.
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
Scope
The Atlas of Genetics and Cytogenetics in Oncology and Haematology is a peer reviewed on-line journal in
open access, devoted to genes, cytogenetics, and clinical entities in cancer, and cancer-prone diseases.
It presents structured review articles ("cards") on genes, leukaemias, solid tumours, cancer-prone diseases, more
traditional review articles on these and also on surrounding topics ("deep insights"), case reports in hematology,
and educational items in the various related topics for students in Medicine and in Sciences.
Editorial correspondance
Jean-Loup Huret Genetics, Department of Medical Information,
University Hospital
F-86021 Poitiers, France
tel +33 5 49 44 45 46 or +33 5 49 45 47 67
jlhuret@AtlasGeneticsOncology.org or Editorial@AtlasGeneticsOncology.org
Staff Mohammad Ahmad, Mélanie Arsaban, Houa Delabrousse, Marie-Christine Jacquemot-Perbal, Maureen
Labarussias, Vanessa Le Berre, Anne Malo, Catherine Morel-Pair, Laurent Rassinoux, Sylvie Yau Chun Wan -
Senon, Alain Zasadzinski.
Philippe Dessen is the Database Director, and Alain Bernheim the Chairman of the on-line version (Gustave
Roussy Institute – Villejuif – France).
The Atlas of Genetics and Cytogenetics in Oncology and Haematology (ISSN 1768-3262) is published 12 times
a year by ARMGHM, a non profit organisation, and by the INstitute for Scientific and Technical Information of
the French National Center for Scientific Research (INIST-CNRS) since 2008.
The Atlas is hosted by INIST-CNRS (http://www.inist.fr)
http://AtlasGeneticsOncology.org
© ATLAS - ISSN 1768-3262
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1)
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
Editor
Jean-Loup Huret
(Poitiers, France)
Editorial Board
Sreeparna Banerjee (Ankara, Turkey) Solid Tumours Section
Alessandro Beghini (Milan, Italy) Genes Section
Anne von Bergh (Rotterdam, The Netherlands) Genes / Leukaemia Sections
Judith Bovée (Leiden, The Netherlands) Solid Tumours Section
Vasantha Brito-Babapulle (London, UK) Leukaemia Section
Charles Buys (Groningen, The Netherlands) Deep Insights Section
Anne Marie Capodano (Marseille, France) Solid Tumours Section
Fei Chen (Morgantown, West Virginia) Genes / Deep Insights Sections
Antonio Cuneo (Ferrara, Italy) Leukaemia Section
Paola Dal Cin (Boston, Massachussetts) Genes / Solid Tumours Section
Louis Dallaire (Montreal, Canada) Education Section
Brigitte Debuire (Villejuif, France) Deep Insights Section
François Desangles (Paris, France) Leukaemia / Solid Tumours Sections
Enric Domingo-Villanueva (London, UK) Solid Tumours Section
Ayse Erson (Ankara, Turkey) Solid Tumours Section
Richard Gatti (Los Angeles, California) Cancer-Prone Diseases / Deep Insights Sections
Ad Geurts van Kessel (Nijmegen, The Netherlands) Cancer-Prone Diseases Section
Oskar Haas (Vienna, Austria) Genes / Leukaemia Sections
Anne Hagemeijer (Leuven, Belgium) Deep Insights Section
Nyla Heerema (Colombus, Ohio) Leukaemia Section
Jim Heighway (Liverpool, UK) Genes / Deep Insights Sections
Sakari Knuutila (Helsinki, Finland) Deep Insights Section
Lidia Larizza (Milano, Italy) Solid Tumours Section
Lisa Lee-Jones (Newcastle, UK) Solid Tumours Section
Edmond Ma (Hong Kong, China) Leukaemia Section
Roderick McLeod (Braunschweig, Germany) Deep Insights / Education Sections
Cristina Mecucci (Perugia, Italy) Genes / Leukaemia Sections
Yasmin Mehraein (Homburg, Germany) Cancer-Prone Diseases Section
Fredrik Mertens (Lund, Sweden) Solid Tumours Section
Konstantin Miller (Hannover, Germany) Education Section
Felix Mitelman (Lund, Sweden) Deep Insights Section
Hossain Mossafa (Cergy Pontoise, France) Leukaemia Section
Stefan Nagel (Braunschweig, Germany) Deep Insights / Education Sections
Florence Pedeutour (Nice, France) Genes / Solid Tumours Sections
Elizabeth Petty (Ann Harbor, Michigan) Deep Insights Section
Susana Raimondi (Memphis, Tennesse) Genes / Leukaemia Section
Mariano Rocchi (Bari, Italy) Genes Section
Alain Sarasin (Villejuif, France) Cancer-Prone Diseases Section
Albert Schinzel (Schwerzenbach, Switzerland) Education Section
Clelia Storlazzi (Bari, Italy) Genes Section
Sabine Strehl (Vienna, Austria) Genes / Leukaemia Sections
Nancy Uhrhammer (Clermont Ferrand, France) Genes / Cancer-Prone Diseases Sections
Dan Van Dyke (Rochester, Minnesota) Education Section
Roberta Vanni (Montserrato, Italy) Solid Tumours Section
Franck Viguié (Paris, France) Leukaemia Section
José Luis Vizmanos (Pamplona, Spain) Leukaemia Section
Thomas Wan (Hong Kong, China) Genes / Leukaemia Sections
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1)
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
Volume 15, Number 1, January 2011
Table of contents
Gene Section
CCR1 (chemokine (C-C motif) receptor 1) 1 Qiang Gao, Jia Fan
GSK3B (glycogen synthase kinase 3 beta) 7 Dinesh Kumar Thotala, Eugenia M Yazlovitskaya
MAPKAPK2 (mitogen-activated protein kinase-activated protein kinase 2) 11 Roberta Felix, Veruska Alves, Andre Vettore, Gisele Colleoni
MINA (MYC induced nuclear antigen) 15 Makoto Tsuneoka, Kengo Okamoto, Yuji Tanaka
NKX2-1 (NK2 homeobox 1) 19 Theresia Wilbertz, Sebastian Maier, Sven Perner
NUDT6 (nudix (nucleoside diphosphate linked moiety X)-type motif 6) 29 Leigh-Ann MacFarlane, Paul Murphy
PARVB (parvin, beta) 34 Cameron N Johnstone
PIAS3 (protein inhibitor of activated STAT, 3) 38 Gilles Spoden, Werner Zwerschke
PSEN2 (presenilin 2 (Alzheimer disease 4)) 42 Morgan Newman
RASSF6 (Ras association (RalGDS/AF-6) domain family member 6) 46 Luke B Hesson, Farida Latif
RPA2 (replication protein A2, 32kDa) 54 Anar KZ Murphy, James A Borowiec
S100A7 (S100 calcium binding protein A7) 58 Jill I Murray, Martin J Boulanger
SOX10 (SRY (sex determining region Y)-box 10) 64 Michael Wegner
TNFSF18 (tumor necrosis factor (ligand) superfamily, member 18) 67 Theresa Placke, Hans-Georg Kopp, Benjamin Joachim Schmiedel, Helmut Rainer Salih
USF1 (upstream transcription factor 1) 72 Adrie JM Verhoeven
WNK2 (WNK lysine deficient protein kinase 2) 76 Peter Jordan
Leukaemia Section
t(1;2)(p36;p21) 79 Jean-Loup Huret
t(11;14)(q13;q32) in multiple myeloma Huret JL, Laï JL
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1)
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
t(2;18)(q11;q21) 81 Jean-Loup Huret
t(2;21)(q11;q22) 83 Jean-Loup Huret
Solid Tumour Section
Liver: Nested stromal epithelial tumor 85 Y Albert Yeh
Deep Insight Section
Inhibitors of the heat shock protein 90: from cancer clinical trials to neurodegenerative diseases 88 Jean François Peyrat, Samir Messaoudi, Jean Daniel Brion, Mouad Alami
LDI-PCR in Cancer Translocation Mapping 105 Björn Schneider, Hans G Drexler, Roderick AF MacLeod
Case Report Section
Unbalanced rearrangement der(9;18)(p10;q10) in a patient with polycythemia vera 114 Xinjie Xu, Xueyan Chen, Elizabeth A Rauch, Eric B Johnson, Kate J Thompson, Jennifer JS Laffin,
Gordana Raca, Daniel F Kurtycz
Gene Section Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 1
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
CCR1 (chemokine (C-C motif) receptor 1) Qiang Gao, Jia Fan
Liver Cancer Institute, Zhong Shan Hospital and Shanghai Medical School, Fudan University,
Shanghai, P R China (QG, JF)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/CCR1ID44379ch3p21.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI CCR1ID44379ch3p21.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity Other names: CD191, CKR-1, CKR1, CMKBR1,
HM145, MIP1aR, SCYAR1
HGNC (Hugo): CCR1
Location: 3p21.31
DNA/RNA
Note
CCR1, a member of the beta chemokine receptor
family, is a seven transmembrane protein similar to
G protein-coupled receptors. CCR1 is the first
human CC chemokine receptor to be identified at
the cDNA level. It has a functional viral homolog,
US28, which is a human cytomegalovirus.The
ligands of this receptor include macrophage
inflammatory protein 1 alpha (MIP-1 alpha),
regulated on activation normal T expressed and
secreted protein (RANTES), monocyte
chemoattractant protein 3 (MCP-3), and myeloid
progenitor inhibitory factor-1 (MPIF-1). This gene
and other chemokine receptor genes, including
CCR2, CCRL2, CCR3, CCR5 and CCXCR1, form
a gene cluster on chromosome 3p.
Description
Sequence length: 6633 bp; coding sequence: CDS
72-1139. 2 exons; number of SNPs: 97.
Transcription
2690 bp mRNA, no alternative splicing.
Pseudogene
No pseudogenes have been reported for CCR1.
Protein
Note
Chemokine receptors are cytokine receptors found
on the surface of certain cells, which interact with a
type of cytokine called a chemokine. They each
have a 7 transmembrane structure and couple to G-
protein for signal transduction within a cell, making
them members of a large protein family of G
protein-coupled receptors. Following interaction
with their specific chemokine ligands, chemokine
receptors trigger a flux in intracellular calcium
(Ca2+
) ions (calcium signaling). This causes cell
responses, including the onset of a process known
as chemotaxis that traffics the cell to a desired
location within the organism.
CCR1 (chemokine (C-C motif) receptor 1) Gao Q, Fan J
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 2
Predicted structure and amino acid sequence of CCR1. The typical serpentine structure is depicted with three extracellular
(top) and three intracellular (bottom) loops and seven transmembrane domains. The shaded horizontal band represents the cell membrane. Amino acids are listed with a single letter code.
Chemokine receptors share many common
structural features; they are composed of about 350
amino acids that are divided into a short and acidic
N-terminal end, seven helical transmembrane
domains with three intracellular and three
extracellular hydrophilic loops, and an intracellular
C-terminus containing serine and threonine residues
that act as phosphorylation sites during receptor
regulation. The first two extracellular loops of
chemokine receptors are linked together by
disulfide bonding between two conserved cysteine
residues. The N-terminal end of a chemokine
receptor binds to chemokine(s) and is important for
ligand specificity. G-proteins couple to the C-
terminal end, which is important for receptor
signaling following ligand binding.
Description
355 amino acids; 41173 Da.
Expression
Monocyte/macrophages; T cells; platelets; tonsil B
lymphocytes; blood derived mast cells, dendritic
cells, basophils and eosinophils; bone marrow
stromal cells; microvascular endothelial cells;
vascular smooth muscle cells.
Localisation
Cell membrane; multi-pass membrane protein.
Function
Receptor for a C-C type chemokine. Binds to CCL3
(MIP-1-alpha), CCL5 (RANTES), CCL7 (MCP-3),
CCL9 (MIP-1-gamma), CCL14 (HCC-1), CCL15
(MIP-1-delta), CCL16 (HCC-4) and CCL23 (MIP-
3), and, less efficiently, to MIP-1-beta or MCP-1
and subsequently transduces a signal by increasing
the intracellular calcium ions level. The major
function of CCR1 is to regulate leukocyte
trafficking in hematopoiesis and in innate and
adaptive immunity. Other functions include
angiogenic activity, ischemia/reperfusion injury,
immune-cell differentiation, phagocyte activation,
and affecting stem cell proliferation.
Homology
CCR1 protein contains considerable amino acid
sequence homology to other C-C chemokines:
CCR2B (56%), CCR3 (54%), CCR4 (49%), CCR5
(55%).
Implicated in
Hematolymphoid neoplasia
Prognosis
CCR1 expression correlates with overall survival in
the non-germinal center subtype of diffuse large B-
CCR1 (chemokine (C-C motif) receptor 1) Gao Q, Fan J
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 3
cell lymphoma. In follicular lymphoma, high levels
of CCR1 are associated with a shorter survival
interval, and CCR1 is a marker of an immune
switch between macrophages and a T cell-dominant
response.
Oncogenesis
CCR1 is expressed in intraepithelial B cells of
human tonsil and granulocytic/monocytic cells in
the bone marrow. Immunohistochemical analysis of
944 cases of hematolymphoid neoplasia identified
CCR1 expression in a subset of B- and T-cell
lymphomas, plasma cell myeloma, acute myeloid
leukemia, and classical Hodgkin lymphoma. In 13
patients with chronic lymphocytic leukemia (CLL),
9 with hairy cell leukemia (HCL), 5 with mantle
cell lymphoma (MCL), 5 with marginal zone B-cell
lymphoma (MZL), 6 with small lymphocytic
lymphoma (SLL), and 5 with follicular cell
lymphoma (FCL), flow cytometry analysis
demonstrated that CCR1 was expressed in 70% of
patients with CLL and 40% of those with HCL but
was lacking in patients with MCL, MZL, SLL, and
circulating normal B cells.
Circulating CD3+ T cells derived from healthy
individuals and acute myelogenous leukemia
patients with therapy-induced cytopenia after
conventional chemotherapy or allogeneic stem cell
transplantation showed no qualitative differences in
CCR1 expression, that is, low expression for all the
three groups.
Multiple myeloma
Prognosis
In 80 multiple myeloma (MM) patients with bone
marrow samples, patients with active disease
showed a significantly lower expression of CCR1,
CCR2, as well as CXCR4 than patients with non-
active disease. This chemokine receptor expression
profile correlated with serum beta2-microglobulin,
C-reactive protein and hemoglobin. Multivariate
analysis identified the chemokine receptor
expression profile as an independent prognostic
factor.
Oncogenesis
Human MM cells express at least three different
chemokine receptors that are functionally involved
in MM cell migration, i.e. CCR1, CCR2 and
CXCR4, some also CCR6 and CXCR3. cDNA
arrays identified CCR1 and CCR2 are
overexpressed in myeloma cells compared to
autologous B-lymphoblastoid cell lines. The
expression of CCR1 and the migration to their
ligands, RANTES and MIP-1alpha, respectively,
were demonstrated in MM cell lines and primary
MM cells.
Osteoclasts (OCL) secrete high levels of CCL3 and
MM cells the express CCR1, the interaction
between which plays a key role in the pathogenesis
of MM-related osteolytic bone disease. Through
CCL3-CCR1 axis OCL cells promote OCL
formation and, in turn, OCL enhance MM cell
proliferation.
In murine models of MM, MIP-1alpha, an OCL
stimulatory factor produced by primary MM cells,
increases bone destruction and tumor burden, by
interacting with chemokine receptors CCR1 and
CCR5 that widely expressed in human OCL
precursors, myeloma cell lines, and purified
marrow plasma cells from MM patients.
Neutralizing antibodies to CCR1 or CCR5 inhibited
MIP-1alpha-induced OCL formation. Furthermore,
MCP-3, which binds CCR1 but not CCR5, and the
CCR1-specific antagonist, BX471, markedly
inhibited OCL formation stimulated with MIP-
1alpha. Anti-CCR1, anti-CCR5, or BX471 also
inhibited the upregulation of beta1 integrin
myeloma cells induced by MIP-1alpha, as well as
the adherence of myeloma cells to stromal cells and
IL-6 production by stromal cells in response to
myeloma cells.
The oncogene c-maf is translocated in
approximately 5%-10% of MM. By gene
expression profiling, three c-maf target genes,
cyclin D2, integrin beta7 and CCR1, were
identified.
Hepatocellular carcinoma
Oncogenesis
Hepatic myofibroblast LI90 cells express and
secrete MCP-1/CCL2. Through its receptors CCR1
and CCR2 as well, LI90 induces human
hepatocellular carcinoma (HCC) Huh7 cell
migration and invasion, which are strongly
inhibited by heparin, beta-D-xyloside and anti-
syndecan-1 and -4 antibodies. RANTES/CCL5
strongly stimulates the migration and the invasion
of Huh7 cells by stimulating the tyrosine
phosphorylation of focal adhesion kinase as well as
activating matrix metalloproteinase-9, and to a
lesser extent that of Hep3B cells. The RANTES-
induced migration and invasion of Huh7 cells are
also strongly inhibited by anti-CCR1 antibodies and
heparin, as well as by beta-d-xyloside treatment of
the cells, suggesting that CCR1 and
glycosaminoglycans are involved in these events.
We found that the miRNA-mediated knockdown
expression of CCR1 significantly inhibited the
invasive ability of and reduced the secretion of
MMP-2 in hepatocellular carcinoma HCCLM3
cells, but only had a minor effect on the cellular
proliferation. CCR1 expression was also detected
on primary HCC cells and to a lesser degree, on
endothelial cells in HCC tissues but not in normal
liver tissues. Similarly, CCL3 expression was
detected in HCC cells, endothelial cells, and to a
lesser degree, fibroblast-like cells in HCC tissue,
whereas only occasional vascular endothelial cells
and inflammatory cells in normal liver tissues were
weakly positive for CCL3. IL-1 enhances the local
production of CCL3, which interact with CCR1
expressed on HCC cells, in an autocrine and/or
CCR1 (chemokine (C-C motif) receptor 1) Gao Q, Fan J
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 4
paracrine manner. In a murine HCC model, injected
tumor cells were transfected with HSV-thymidine
kinase gene and then treated with ganciclovir
(GCV). GCV treatment induced massive tumor cell
apoptosis accompanied with intratumoral
CCR1+CCR5+ dendritic cell infiltration. Tumor-
infiltrating T cells and macrophages expressed
CCL3, suggesting CCR1-CCL3 play a crucial role
in the regulation of intratumoral dendritic cell
accumulation and the subsequent establishment of
tumor immunity following induction of tumor
apoptosis by suicide genes. CCL3 and CCR1 are
also expressed in 2 different models of HCC, i.e.,
N-nitrosodiethylamine (DEN)-induced HCC and
HCC induced by hepatitis B virus. After DEN
treatment, tumor foci number and sizes were
remarkably reduced in CCR1- and CCL3-deficient
mice, comparing with wild-type (WT) mice. Also,
tumor angiogenesis markedly diminished,
intratumoral Kupffer cells number reduced, MMP9
gene expression attenuated and MMP9+ cell
numbers decreased in CCL3- and CCR1-deficient
mice, as compared with WT mice. These
observations suggest the contribution of the CCR1-
CCL3 axis to HCC progression.
Colorectal cancer
Prognosis
The expression of CCR1 is higher in colorectal
carcinoma than normal tissues, and correlates with
lymph node metastasis, deep invasion, poor
differentiation and advanced Dukes' stage.
Oncogenesis
Inactivation of TGF-beta family signaling within
colon cancer increases CCL9 and promotes
recruitment of the matrix metalloproteinase-
expressing stromal cells that carry CCR1. Lack of
CCR1 prevents the accumulation of MMP-
expressing cells at the invasive front and suppresses
tumor invasion. In a murine model of invasive
colorectal cancer in which TGF-beta family
signaling is blocked, CD34+ CCR1+ immature
myeloid cell is recruited from the bone marrow to
the tumor invasive front where expression of CCL9
is increased. These immature myeloid cells express
MMP9, MMP2 and CCR1 and migrate toward the
ligand CCL9. Lack of CCR1 prevents accumulation
of CD34+ immature myeloid cell at the invasive
front and suppresses tumor invasion.
Non-small cell lung cancer
Oncogenesis
CCR1 expression correlated with the aggressive
phenotype of the non-small cell lung cancer
(NSCLC) cells. CCR1 knockdown significantly
suppressed the invasiveness of NSCLC cells and
significantly reduced the expression of matrix
metalloproteinase-9, but had only a minor effect on
cell proliferation.
Oral squamous cell carcinoma
Oncogenesis
Expression of CCL3 and CCR1 is significant higher
in oral squamous cell carcinoma compared with the
normal controls. The percentages of CCL3+ and
CCR1+ cells were observed to be similar in
parenchyma and stroma in cases without lymph
node metastasis when compared with lymph node
metastasis positive cases.
Ovarian cancer
Oncogenesis
mRNA for CCR1, -2a, -2b, -3, -4, -5, and -8 was
detected in cells from human ovarian cancer ascites.
Further, flowcytometry showed CD14+
macrophages within ascites consistently expressed
CCR1, -2, and -5, and >60% of all T cells
expressed CCR1. Although ovarian cancer ascitic
and blood monocyte/macrophages express CCR1,
they failed to migrate in response to the RANTES.
Compared with that of normal blood, cell surface
expression level for CCR1 was higher in ascites. In
a monocytic cell line in vitro, CCR1 mRNA
expression was increased 5-fold by hypoxia. In 25
patients with ovarian cancer, CCR1 was detected in
samples from 75% of patients, where CCR1
localised to macrophages and lymphocytes, and
there was a correlation between numbers of CD8+
cells and CCR1+ cells.
Prostate cancer
Oncogenesis
Androgen receptor negative human prostate cancer
cell line DU-145 cells selectively expressed
CXCR4 and CCR1 at high levels compared with
DU-145/AR cells that express androgen receptor.
DU-145 showed vigorous migratory responses to
CXCL12 and CCL3. In contrast, neither CXCL12
nor CCL3 affected the migration of DU-145/AR
cells.
Breast cancer
Oncogenesis
The expression of CCR5 was higher than that of
CCR1 in the peripheral blood mononuclear cells
(PBMC) of healthy women, while the PBMC of the
breast cancer patients showed overexpression of
CCR1 and downregulation of CCR5. The
differential effects of MIP-1alpha and MIP-1beta
on the PBMC of healthy women and breast cancer
patients correlated with the expression levels of
CCR1 and CCR5 in these monocytes. In murine
model of breast cancer, CCL5 (RANTES) was
produced by the tumor cells, and its receptors,
CCR1 and CCR5, were expressed by the infiltrating
leukocytes. In mice treatment with Met-CCL5, an
antagonist of CCR1 and CCR5, the volume and
weight of tumors were significantly decreased
compared with control-treated tumors.
CCR1 (chemokine (C-C motif) receptor 1) Gao Q, Fan J
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 5
The total cell number obtained after collagenase
digestion was decreased in Met-CCL5-treated
tumors as was the proportion of infiltrating
macrophages. Furthermore, chemokine antagonist
treatment increased stromal development and
necrosis.
Glioma
Oncogenesis
Co-cultured human glioma U87 cells induced an
activated phenotype in HUVECs. These tumour-
activated endothelial cells coordinately expressed
matching pairs of receptors/ligands were found to
be, including CCR1-RANTES axis.
Osteogenic sarcoma
Oncogenesis
The activities of phospholipase C (PLC), protein
kinase C delta (PKCdelta) and NF-kappaB were
enhanced by Lkn-1 (CCL15) stimulation on CCR1+
human osteogenic sarcoma cells. Inhibitors of G
protein, PLC, PKCdelta and NF-kappaB inhibited
the chemotactic activity of Lkn-1 on CCR1+
osteogenic sarcoma cells indicating that Lkn-1-
induced chemotaxis involving these signaling
pathways.
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Ribeiro S, Horuk R. The clinical potential of chemokine receptor antagonists. Pharmacol Ther. 2005 Jul;107(1):44-58
Akashi T, Koizumi K, Nagakawa O, Fuse H, Saiki I. Androgen receptor negatively influences the expression of chemokine receptors (CXCR4, CCR1) and ligand-mediated migration in prostate cancer DU-145. Oncol Rep. 2006 Oct;16(4):831-6
Menu E, De Leenheer E, De Raeve H, Coulton L, Imanishi T, Miyashita K, Van Valckenborgh E, Van Riet I, Van Camp B, Horuk R, Croucher P, Vanderkerken K. Role of CCR1 and CCR5 in homing and growth of multiple myeloma and in the development of osteolytic lesions: a study in the 5TMM model. Clin Exp Metastasis. 2006;23(5-6):291-300
Nath A, Chattopadhya S, Chattopadhyay U, Sharma NK. Macrophage inflammatory protein (MIP)1alpha and MIP1beta differentially regulate release of inflammatory cytokines and generation of tumoricidal monocytes in malignancy. Cancer Immunol Immunother. 2006 Dec;55(12):1534-41
Vande Broek I, Leleu X, Schots R, Facon T, Vanderkerken K, Van Camp B, Van Riet I. Clinical significance of chemokine receptor (CCR1, CCR2 and CXCR4) expression in human myeloma cells: the association with disease activity and survival. Haematologica. 2006 Feb;91(2):200-6
Yang X, Lu P, Fujii C, Nakamoto Y, Gao JL, Kaneko S, Murphy PM, Mukaida N. Essential contribution of a chemokine, CCL3, and its receptor, CCR1, to hepatocellular carcinoma progression. Int J Cancer. 2006 Apr 15;118(8):1869-76
Kitamura T, Kometani K, Hashida H, Matsunaga A, Miyoshi H, Hosogi H, Aoki M, Oshima M, Hattori M, Takabayashi A, Minato N, Taketo MM. SMAD4-deficient intestinal tumors recruit CCR1+ myeloid cells that promote invasion. Nat Genet. 2007 Apr;39(4):467-75
CCR1 (chemokine (C-C motif) receptor 1) Gao Q, Fan J
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Kitamura T, Taketo MM. Keeping out the bad guys: gateway to cellular target therapy. Cancer Res. 2007 Nov 1;67(21):10099-102
Silva TA, Ribeiro FL, Oliveira-Neto HH, Watanabe S, Alencar Rde C, Fukada SY, Cunha FQ, Leles CR, Mendonça EF, Batista AC. Dual role of CCL3/CCR1 in oral squamous cell carcinoma: implications in tumor metastasis and local host defense. Oncol Rep. 2007 Nov;18(5):1107-13
Sutton A, Friand V, Papy-Garcia D, Dagouassat M, Martin L, Vassy R, Haddad O, Sainte-Catherine O, Kraemer M, Saffar L, Perret GY, Courty J, Gattegno L, Charnaux N. Glycosaminoglycans and their synthetic mimetics inhibit RANTES-induced migration and invasion of human hepatoma cells. Mol Cancer Ther. 2007 Nov;6(11):2948-58
Vallet S, Raje N, Ishitsuka K, Hideshima T, Podar K, Chhetri S, Pozzi S, Breitkreutz I, Kiziltepe T, Yasui H, Ocio EM, Shiraishi N, Jin J, Okawa Y, Ikeda H, Mukherjee S, Vaghela N, Cirstea D, Ladetto M, Boccadoro M, Anderson KC. MLN3897, a novel CCR1 inhibitor, impairs osteoclastogenesis and inhibits the interaction of multiple myeloma cells and osteoclasts. Blood. 2007 Nov 15;110(10):3744-52
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cancer. Cancer Immunol Immunother. 2008 May;57(5):635-45
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This article should be referenced as such:
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Gene Section Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 7
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
GSK3B (glycogen synthase kinase 3 beta) Dinesh Kumar Thotala, Eugenia M Yazlovitskaya
Department of Radiation Oncology, Vanderbilt Ingram Cancer Center, Vanderbilt University, SS1411
Medical Center North, 1161 21 Avenue S, Nashville, TN 37232, USA (DKT, EMY)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/GSK3BID40761ch3q13.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI GSK3BID40761ch3q13.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity
Other names: EC 2.7.11.26
HGNC (Hugo): GS3KB
Location: 3q13.33
Local order:
Human: Nuclear receptor subfamily 1, group I,
member 2 (NR1I2); GSK3B; G-protein coupled
receptor 156 (GPR156).
Mouse: G-protein coupled receptor 156 (Gpr156);
Gsk3b; Nuclear receptor subfamily 1, group I,
member 2 (Nr1i2).
DNA/RNA
Description
According to Entrez-Gene, human GSK3B maps to
locus NC_000003.11.
This gene contains 12 exons that encompass
266971 bp of genomic DNA. In mice, GSK3B
maps to NC_000082.5 and contains 11 exons that
span 157079 bp of DNA within the mouse genome.
Transcription
Human GSK3B mRNA (NM_002193.3) consists of
7134 bp, and murine GSK3B mRNA (NM_019827)
contains 8298 bp. Alternatively spliced transcript
variants encoding different isoforms (1 and 2) have
been found for human gene. Transcript variant 2 is
missing an in-frame coding exon (9) compared to
variant 1, resulting in a shorter isoform 2 lacking a
13 aa segment compared to isoform 1.
Pseudogene
No pseudogene has been identified for GSK3B.
A) Human GSK3B gene, isoform 1. B) Mouse Gsk3b gene. GSK3B is comprised of 12 exons in human and 11 exons in
mouse. The ATG start codon is located within exon 1 and the TAG stop codon is found in exon 12 (Human) and 11 (Mouse). The sizes of exons for human gene 1-12 are 1071 bp, 191 bp, 85 bp, 110 bp, 130 bp, 106 bp, 97 bp, 95 bp, 38 bp, 186 bp, 98 bp and 604 bp, respectively. The sizes of exons for mouse gene 1-11 are 1613 bp, 193 bp, 83 bp, 110 bp, 130 bp, 106 bp, 97 bp, 95 bp,
186 bp, 98 bp and 5577 bp, respectively.
GSK3B (glycogen synthase kinase 3 beta) Thotala DK, Yazlovitskaya EM
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 8
GSK3B structure. GSK3B is a 46-47 kDa protein consisting of 433 and 420 amino acids in human and mouse respectively. The
protein contains an N-terminal domain, a kinase domain and a C-terminal domain. Phosphorylation of Tyr216 located in the T-loop (activation site) facilitates substrate phosphorylation by GSK3B but is not strictly required for its kinase activity.
Phosphorylation of GSK3B at Ser9 in N-terminal region leads to inhibition of its kinase activity. Binding domain (BD) includes GSK3B specific binding sites for substrates and protein complexes (e.g., p53).
Protein
Description
Glycogen synthase kinase-3 beta (GSK3B) was
named due to its ability to phosphorylate and
inactivate glycogen synthase. GSK3B is a
multifunctional serine/threonine kinase which has
been implicated in multiple biological processes
including embryonic development, cell
differentiation, apoptosis and insulin response.
GSK3B is a key component in neuronal functions
and has been implicated in major diseases involving
the central nervous system.
Expression
GSK3B was originally isolated from the skeletal
muscle but it is ubiquitously expressed in almost all
the tissues. However, abundant expression is
detected in brain tissue, especially in the neurons
when compared to the astrocytes. The high level of
expression in the brain is due to its vital role in the
neuronal signaling. Dysregulation of GSK3B
expression leads to various pathological conditions
such as diabetes or insulin resistance, neuronal
dysfunction and neuronal diseases.
Localisation
GSK3B is generally considered a cytosolic protein;
however, it is reported to be present in the nucleus
and mitochondria. Nuclear and mitochondrial
localization of GSK3B correlates with its higher
kinase activity compared to cytosolic protein.
Translocation and specific cellular localization of
GSK3B determine its involvement in signaling
pathways, regulate its interaction with substrates
and participation in protein complex formation, and
influence gene expression and transcription.
Function
GSK3B is a multifunctional protein kinase which is
implicated in a large number of cellular processes
and diseases. GSK3B is regulated by serine
(inhibitory) and tyrosine (activating)
phosphorylation. More than 40 proteins have been
reported to be phosphorylated by GSK3B. GSK3B
substrates include metabolic and signaling proteins
like glycogen synthase, Acetyl CoA carboxylase,
Axin, Cyclin D1; structural proteins like Tau,
neural cell adhesion protein (NCAM); transcription
factors like beta-catenin, p53, Myc, NFkappaB,
CREB and AP-1; apoptotic-related proteins like
Bax and p53. GSK3B also regulates various cellular
processes by binding to protein complexes.
Homology
The GSK3B gene is conserved in human,
chimpanzee, dog, cow, rat, chicken, zebrafish, fruit
fly, mosquito, C. elegans, A. thaliana, rice, and P.
falciparum.
Mutations
Germinal
1. Several rare sequence variants in GSK3B were
identified in the case-control study of patients with
probable Alzheimer disease (AD), familial
frontotemporal dementia (FTD), primary
progressive aphasia, and aged healthy subjects. An
intronic polymorphism (IVS2-68G>A) occurred at
more than twice the frequency among patients with
FTD (10.8%) and patients with AD (14.6%) than in
aged healthy subjects (4.1%).
2. GSK3beta promoter single-nucleotide
polymorphism (rs334558) influences transcriptional
strength, and the less active form was associated
with less detrimental clinical features of mood
disorders. Effect of rs334558 was studied on grey
matter volumes of patients affected by chronic
schizophrenia. Carriers of the less active C allele
variant showed significantly higher brain volumes
in an area encompassing posterior regions of right
middle and superior temporal gyrus, within the
boundaries of Brodmann area 21. The temporal
lobe is the brain parenchymal region with the most
consistently documented morphometric
abnormalities in schizophrenia, and
neuropathological processes in these regions
develop soon at the beginning of the illness.
Implicated in
Ovarian cancer
Note
Ovarian cancer is a leading cause of death from
gynecological malignancies. GSK3B promotes
ovarian cancer cell proliferation by regulating
GSK3B (glycogen synthase kinase 3 beta) Thotala DK, Yazlovitskaya EM
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 9
Cyclin D1. GSK3B-dependent increased Cyclin D1
expression in ovarian cancer cells supports a
possibility that GSK3B is involved in ovarian
tumor chemotherapy resistance. Therefore, it is
possible that combination of traditional
chemotherapy and GSK3B inhibitors would benefit
ovarian cancer patient response.
Prostate cancer
Note
Androgen receptor (AR) regulates growth of
normal and cancer prostate cells. AR
phosphorylation status is associated with its
transcriptional activation. GSK3B interacts directly
with the AR, modulates AR signaling and plays
important role in the control of the proliferation of
normal and malignant androgen-regulated tissues.
Therefore, pharmacological inhibitors designed to
increase GSK3B activity could be useful in prostate
cancer therapy.
Pancreatic cancer
Note
It was shown that pancreatic cancer cells contain a
pool of active GSK3B, and that pharmacological
inhibition of GSK3B kinase activity using small
molecule inhibitors or genetic depletion of GSK3B
by RNA interference leads to decreased cancer cell
proliferation and survival. Hence GSK3B has
potential as an important new target in the treatment
of pancreatic cancer.
Colorectal cancer
Note
Colon cancer cell lines and colon cells from
colorectal cancer patients have higher levels of
GSK3B expression than their normal counterparts.
Inhibition of GSK3B activity either by chemical
inhibitors or by expression by RNA interference
targeting GSK3B induced apoptosis and attenuation
of proliferation of colon cancer cells in vitro. Hence
GSK3B has a potential as therapeutic target in
colorectal cancer.
Neuroblastoma
Note
Treatment of B65 neuroblastoma cell line with
GSK3B inhibitors Lithium or SB415286 caused a
decrease in cell proliferation that was associated
with G2/M cell cycle arrest due to regulating the
phosphorylation of Cdc2. Therefore, GSK3B and
Cdc2 could be potential pharmacological targets in
neuroblastoma.
Glioblastoma
Note
Glioblastoma is the most frequent malignant tumor
of the brain and represents a subset of cancers that
is mostly nonresponsive to currently available
anticancer treatments.
The current standard therapy for newly diagnosed
glioblastoma consists of surgical resection of the
tumor to the extent that is safe and feasible,
followed by chemotherapy and irradiation. There
has been an emerging paradigm for the combination
of chemotherapy and molecular targeted therapy to
improve therapeutic efficiency. Glioblastoma cells
depend on deregulated GSK3B to survive,
proliferate, and resist chemotherapy and radiation.
Pretreatment with low-dose GSK3B inhibitor
enhanced the cytocidal effect of ionizing radiation
in glioblastoma cells. At the same time, GSK3B
inhibitors have been shown to protect normal
hippocampal neurons from radiation-induced
apoptosis. Therefore, GSK3B inhibition provides
dual benefits for the glioblastoma patients treated
with radiation: by attenuating tumor proliferation
and by protecting host brain tissue from
degradation and allowing its repair.
Insulin resistance and diabetes
Note
Insulin resistance is caused by the inability of
insulin sensitive tissues to respond to insulin and
efficiently clear blood glucose. Insulin signaling
involves autophosphorylation of the insulin
receptor leading to the activation of PI3K which
activates PKB (Akt). The activated PKB
phosphorylates and inactivates GSK3B.
Dysregulation of GSK3B results in impaired insulin
signaling leading to diabetes. Inhibitors of GSK3B
improve insulin signaling and maintain proper
glucose levels.
Alzheimer's disease
Note
Alzheimer's disease (AD) is a chronic disorder that
slowly destroys neurons and causes serious
cognitive disability. The two neuropathologiocal
features of Alzheimer's disease are neurofibrillary
tangles and amyloid plaques. GSK3B has been
implicated in both neuropathologies. In addition,
presenilin 1 (PS1) have been linked to Alzheimer's
disease. Presenilin 1 binds to and regulates GSK3B
activity. Presenilin 1 mutations might compromise
neuronal function by increasing GSK3B activity.
Schizophrenia
Note
Schizophrenia is a severe brain illness in which the
disrupted in schizophrenia 1 (DISC1) gene is
disrupted by a balanced chromosomal translocation.
DISC1 is highly expressed in neural progenitor
cells and required for embryonic brain
development. DISC1 regulates beta-catenin
turnover by inhibiting GSK3B activity. GSK3B
inhibitors are able to normalize progenitor
proliferation and behavioral defects caused by
DISC1 loss of function.
GSK3B (glycogen synthase kinase 3 beta) Thotala DK, Yazlovitskaya EM
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 10
Bipolar affective disorder
Note
Patients with bipolar affective disorder have a
history of experiencing manic episodes that are
often interspersed with depression, and major
depression is commonly referred to as mood
disorders. Lithium, a known GSK3B inhibitor, is
one of the most widely used mood-stabilizing
agents for the treatment of bipolar disorder.
References Lau KF, Miller CC, Anderton BH, Shaw PC. Molecular cloning and characterization of the human glycogen synthase kinase-3beta promoter. Genomics. 1999 Sep 1;60(2):121-8
Dajani R, Fraser E, Roe SM, Young N, Good V, Dale TC, Pearl LH. Crystal structure of glycogen synthase kinase 3 beta: structural basis for phosphate-primed substrate specificity and autoinhibition. Cell. 2001 Jun 15;105(6):721-32
Martinez A, Castro A, Dorronsoro I, Alonso M. Glycogen synthase kinase 3 (GSK-3) inhibitors as new promising drugs for diabetes, neurodegeneration, cancer, and inflammation. Med Res Rev. 2002 Jul;22(4):373-84
Bhat RV, Budd Haeberlein SL, Avila J. Glycogen synthase kinase 3: a drug target for CNS therapies. J Neurochem. 2004 Jun;89(6):1313-7
Cohen P, Goedert M. GSK3 inhibitors: development and therapeutic potential. Nat Rev Drug Discov. 2004 Jun;3(6):479-87
Jope RS, Johnson GV. The glamour and gloom of glycogen synthase kinase-3. Trends Biochem Sci. 2004 Feb;29(2):95-102
Meijer L, Flajolet M, Greengard P. Pharmacological inhibitors of glycogen synthase kinase 3. Trends Pharmacol Sci. 2004 Sep;25(9):471-80
Wang L, Lin HK, Hu YC, Xie S, Yang L, Chang C. Suppression of androgen receptor-mediated transactivation and cell growth by the glycogen synthase kinase 3 beta in prostate cells. J Biol Chem. 2004 Jul 30;279(31):32444-52
Ougolkov AV, Fernandez-Zapico ME, Savoy DN, Urrutia RA, Billadeau DD. Glycogen synthase kinase-3beta participates in nuclear factor kappaB-mediated gene transcription and cell survival in pancreatic cancer cells. Cancer Res. 2005 Mar 15;65(6):2076-81
Shakoori A, Ougolkov A, Yu ZW, Zhang B, Modarressi MH, Billadeau DD, Mai M, Takahashi Y, Minamoto T. Deregulated GSK3beta activity in colorectal cancer: its association with tumor cell survival and proliferation. Biochem Biophys Res Commun. 2005 Sep 9;334(4):1365-73
Cao Q, Lu X, Feng YJ. Glycogen synthase kinase-3beta positively regulates the proliferation of human ovarian cancer cells. Cell Res. 2006 Jul;16(7):671-7
Yazlovitskaya EM, Edwards E, Thotala D, Fu A, Osusky KL, Whetsell WO Jr, Boone B, Shinohara ET, Hallahan DE. Lithium treatment prevents neurocognitive deficit
resulting from cranial irradiation. Cancer Res. 2006 Dec 1;66(23):11179-86
Garcea G, Manson MM, Neal CP, Pattenden CJ, Sutton CD, Dennison AR, Berry DP. Glycogen synthase kinase-3 beta; a new target in pancreatic cancer? Curr Cancer Drug Targets. 2007 May;7(3):209-15
Schaffer BA, Bertram L, Miller BL, Mullin K, Weintraub S, Johnson N, Bigio EH, Mesulam M, Wiedau-Pazos M, Jackson GR, Cummings JL, Cantor RM, Levey AI, Tanzi RE, Geschwind DH. Association of GSK3B with Alzheimer disease and frontotemporal dementia. Arch Neurol. 2008 Oct;65(10):1368-74
Thotala DK, Hallahan DE, Yazlovitskaya EM. Inhibition of glycogen synthase kinase 3 beta attenuates neurocognitive dysfunction resulting from cranial irradiation. Cancer Res. 2008 Jul 15;68(14):5859-68
Eom TY, Jope RS. GSK3 beta N-terminus binding to p53 promotes its acetylation. Mol Cancer. 2009 Mar 5;8:14
Machado-Vieira R, Manji HK, Zarate CA Jr. The role of lithium in the treatment of bipolar disorder: convergent evidence for neurotrophic effects as a unifying hypothesis. Bipolar Disord. 2009 Jun;11 Suppl 2:92-109
Mao Y, Ge X, Frank CL, Madison JM, Koehler AN, Doud MK, Tassa C, Berry EM, Soda T, Singh KK, Biechele T, Petryshen TL, Moon RT, Haggarty SJ, Tsai LH. Disrupted in schizophrenia 1 regulates neuronal progenitor proliferation via modulation of GSK3beta/beta-catenin signaling. Cell. 2009 Mar 20;136(6):1017-31
Miyashita K, Kawakami K, Nakada M, Mai W, Shakoori A, Fujisawa H, Hayashi Y, Hamada J, Minamoto T. Potential therapeutic effect of glycogen synthase kinase 3beta inhibition against human glioblastoma. Clin Cancer Res. 2009 Feb 1;15(3):887-97
Pizarro JG, Folch J, Esparza JL, Jordan J, Pallàs M, Camins A. A molecular study of pathways involved in the inhibition of cell proliferation in neuroblastoma B65 cells by the GSK-3 inhibitors lithium and SB-415286. J Cell Mol Med. 2009 Sep;13(9B):3906-17
Takahashi-Yanaga F, Sasaguri T. Drug development targeting the glycogen synthase kinase-3beta (GSK-3beta)-mediated signal transduction pathway: inhibitors of the Wnt/beta-catenin signaling pathway as novel anticancer drugs. J Pharmacol Sci. 2009 Feb;109(2):179-83
Benedetti F, Poletti S, Radaelli D, Bernasconi A, Cavallaro R, Falini A, Lorenzi C, Pirovano A, Dallaspezia S, Locatelli C, Scotti G, Smeraldi E. Temporal lobe grey matter volume in schizophrenia is associated with a genetic polymorphism influencing glycogen synthase kinase 3-beta activity. Genes Brain Behav. 2010 Jun 1;9(4):365-71
Thotala DK, Geng L, Dickey AK, Hallahan DE, Yazlovitskaya EM. A new class of molecular targeted radioprotectors: GSK-3beta inhibitors. Int J Radiat Oncol Biol Phys. 2010 Feb 1;76(2):557-65
This article should be referenced as such:
Thotala DK, Yazlovitskaya EM. GSK3B (glycogen synthase kinase 3 beta). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):7-10.
Gene Section Mini Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 11
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
MAPKAPK2 (mitogen-activated protein kinase-activated protein kinase 2) Roberta Felix, Veruska Alves, Andre Vettore, Gisele Colleoni
Laboratory of Cancer Molecular Biology, Federal University of Sao Paulo UNIFESP/EPM, Sao
Paulo, Brazil (RF, VA, AV, GC)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/MAPKAPK2ID41295ch1q32.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI MAPKAPK2ID41295ch1q32.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity
Other names: MAPKAPK-2, MK2
HGNC (Hugo): MAPKAPK2
Location: 1q32.1
Note: MAPKAPK2 is involved in many cellular
processes including: stress and inflammatory
response, nuclear export, gene expression
regulation and cell proliferation, acting with p38
MAP gene.
DNA/RNA
Note
MAPKAPK2 encodes a member of the Ser/Thr
protein kinase regulated through direct
phosphorylation by p38 MAP kinase. Inhibition of
the p38 MAPK pathway could be a possible target
to inflammatory diseases therapy. Unfortunately,
blocking p38 MAPK activation "in vivo" implies in
high toxicity and it does not have oral
bioavailability. MAPKAPK2/MK2 inhibitors acting
downstream of p38 could be reasonable solutions to
overcome this problem (Duraisamy et al., 2008).
Description
Size 49,338 bases, starts at 204924912 and ends at
204974256 bp from pter with plus strand
orientation.
Transcription
We found some discordant information regarding
MAPKAPK2 splice variants. Kervinen et al. 2006,
described that the human MAPKAPK2 gene
encodes two alternatively spliced transcripts and
also that this gene contains 14 different introns (13
gt-ag, 1 gc-ag). Transcription produces 9 different
mRNAs, 8 alternatively spliced variants and 1
unspliced form. There are 5 probable alternative
promoters and 3 validated alternative
polyadenylation sites. The mRNAs appear to differ
by truncation of the 5' end, presence or absence of 7
cassette exons, overlapping exons with different
boundaries, alternative splicing or retention of 3
introns.
Pseudogene
ATF4C - Cyclic AMP-dependent transcription
factor ATF-4, localized at chromosome 17,
location: 17q25.1.
Protein
Description
MAPKAPK2 has two alternatively spliced
transcripts, encoding 400 and 370 amino acids, with
sequence heterogeneity from Lys-353 to the C-
terminus. Crystal structure shows: 1) an
autoinhibitory domain, consisting of 328-370
residues; 2) a helix-turn-helix structure that
occupies the substrate-binding cleft of the kinase
domain and inhibits kinase function (ter Haar et al.,
2007).
Localisation
The 400-residue MAPKAPK2 (isoform 1) consists
of an N-terminal Pro-rich region, a kinase domain,
an autoinhibitory domains, and C-terminal nuclear
export (NES) and nuclear localization (NLS)
signals. The 1-370 isoform (isoform 2) lacks NES
and NLS, consistent with its presence only in the
MAPKAPK2 (mitogen-activated protein kinase-activated protein kinase 2)
Felix R, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 12
cytoplasm. MAPKAPK2 also phosphorylates
proteins found in both the nucleus (cAMP-response
element-binding protein, or CREB) and cytoplasm
(HSP25/27 and LSP-1) (Kervinen et al., 2006).
Function
MAPKAPK2 is required for both cytokines
production and cell migration (Kotlyarov et al.,
2002).
MAPKAPK2 is activated upon stress by p38
MAPK, which binds C terminus of MAPKAPK2,
leading to subsequently phosphorylation of its
regulatory sites. After activation, MAPKAPK2 is
transferred from nucleus to cytoplasm, and
cotransport p38 to the new localization. In murine
knockout model, MAPKAPK2 blockage leads to a
dramatic reduction of tumor necrosis factor (TNF)
production in response to lipopolysaccharide
(Kotlyarov et al., 2002). One of the major
substrates of MAPKAPK2 is the heat shock protein
HSP27, which stimulates actin polymerization in
order to facilitate recovery from destruction of
cytoskeleton during cellular stresses.
Homology
Isoform 1 and 2 see figure below.
MAPKAPK2 (mitogen-activated protein kinase-activated protein kinase 2)
Felix R, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 13
Mutations
Note
There is one described mutation: position 804 of
mRNA, allele change GCC to GGC. At protein
level, residue change A [Ala] to G [Gly].
Implicated in
Multiple myeloma (MM)
Note
Hideshima et al. 2004 have shown that
overexpression of HSP27 confers resistance to
bortezomib, a proteasome inhibitor currently used
as front line MM therapy, combined with
corticosteroids and immunomodulatory drugs, such
as thalidomide and lenalidomide. Therefore,
overexpression of MAPKAPK2 could be related to
MM resistance to chemotherapy. They
hypothesized that inhibition of MAPKAPK2
activity could augment bortezomib cytotoxicity by
down regulating HSP27. Felix et al. 2009, at their
gene expression studies, supported further
exploitation of this pathway as therapeutic target in
MM, although immunohistochemistry did not show
high frequency of protein expression in MM (21%)
(Felix et al., 2009).
Panel showing bone marrow samples of MM cases: A -
plasma cells nuclear and cytoplasmatic positivity for MAPKAPK2; B - sample negative for MAPKAPK2 (400X).
Bladder cancer
Note
Kumar et al. 2010, showed that overexpression of
the matrix metalloproteinases MMP-2 and MMP-9
have prognostic value in transitional cell carcinoma
of bladder. p38 MAPK modulated MMP-2/9
mRNA expression and MMP-2/9 activity as
mediators of tumor cells invasive capacity.
Therefore, p38 MAPK inhibition blocks MMP-2/9
activities mediated by MAPKAPK2 (Kumar et al.,
2010).
Skin tumor
Note
Using the two-stage chemical carcinogenesis
model, Johansen et al. 2009 studied the effect of
MAPKAPK2-deficiency and TNF-alpha-deficiency
on skin tumor development in mice. Their findings
demonstrate a dual role of MAPKAPK2 in the early
stages of tumor promotion through regulation of
both the inflammatory response and apoptosis of
DNA-damaged cells. These results also identify
MAPKAPK2 as a possible target for skin
carcinoma therapy (Johansen et al., 2009).
Prostate cancer
Note
TGFbeta is an important regulator of cell adhesion
and motility in a variety of cell types. p38 MAP
kinase is necessary for TGFbeta -mediated up-
regulation of matrix metalloproteinase type 2
(MMP-2), as well as TGFbeta -dependent increases
in prostate cell invasion. Xu et al. 2006
demonstrated, after transient transfection, that both
MAPKAPK2 and HSP27 are necessary for
TGFbeta -mediated increases in MMP-2 activity in
any cell type, as well as prostate cancer cells (Xu et
al., 2006).
Alzheimer's disease (AD)
Note
Culbert et al. 2006 suggested that MAPKAPK2
plays a role in neuroinflammatory and
neurodegenerative diseases, such as AD. The
MAPKAPK2 activation and expression were
increased in lipopolysaccharide (LPS) + interferon
gamma-stimulated microglial cells, demonstrating
MAPKAPK2 ability in eliciting a pro-inflammatory
response. Again, MAPKAPK2 pathway can be
considered a target for control of this degenerative
brain disease.
Psoriatic skin
Note
Alterations in this specific signal transduction
pathway may be involved in increased expression
of proinflammatory cytokines in inflammatory
diseases (Johansen et al., 2006). The increased
activation of MAPKAPK2 is responsible for the
elevated TNFalpha protein expression in psoriatic
MAPKAPK2 (mitogen-activated protein kinase-activated protein kinase 2)
Felix R, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 14
skin, making this pathway a potential target in the
treatment of psoriasis (Johansen et al., 2006).
References Kotlyarov A, Yannoni Y, Fritz S, Laass K, Telliez JB, Pitman D, Lin LL, Gaestel M. Distinct cellular functions of MK2. Mol Cell Biol. 2002 Jul;22(13):4827-35
Hideshima T, Podar K, Chauhan D, Ishitsuka K, Mitsiades C, Tai YT, Hamasaki M, Raje N, Hideshima H, Schreiner G, Nguyen AN, Navas T, Munshi NC, Richardson PG, Higgins LS, Anderson KC. p38 MAPK inhibition enhances PS-341 (bortezomib)-induced cytotoxicity against multiple myeloma cells. Oncogene. 2004 Nov 18;23(54):8766-76
Culbert AA, Skaper SD, Howlett DR, Evans NA, Facci L, Soden PE, Seymour ZM, Guillot F, Gaestel M, Richardson JC. MAPK-activated protein kinase 2 deficiency in microglia inhibits pro-inflammatory mediator release and resultant neurotoxicity. Relevance to neuroinflammation in a transgenic mouse model of Alzheimer disease. J Biol Chem. 2006 Aug 18;281(33):23658-67
Johansen C, Funding AT, Otkjaer K, Kragballe K, Jensen UB, Madsen M, Binderup L, Skak-Nielsen T, Fjording MS, Iversen L. Protein expression of TNF-alpha in psoriatic skin is regulated at a posttranscriptional level by MAPK-activated protein kinase 2. J Immunol. 2006 Feb 1;176(3):1431-8
Kervinen J, Ma H, Bayoumy S, Schubert C, Milligan C, Lewandowski F, Moriarty K, Desjarlais RL, Ramachandren K, Wang H, Harris CA, Grasberger B, Todd M, Springer BA, Deckman I. Effect of construct design on MAPKAP kinase-2 activity, thermodynamic stability and ligand-binding affinity. Arch Biochem Biophys. 2006 May 15;449(1-2):47-56
Xu L, Chen S, Bergan RC. MAPKAPK2 and HSP27 are downstream effectors of p38 MAP kinase-mediated matrix metalloproteinase type 2 activation and cell invasion in human prostate cancer. Oncogene. 2006 May 18;25(21):2987-98
ter Haar E, Prabhakar P, Liu X, Lepre C. Crystal structure of the p38 alpha-MAPKAP kinase 2 heterodimer. J Biol Chem. 2007 Mar 30;282(13):9733-9
Duraisamy S, Bajpai M, Bughani U, Dastidar SG, Ray A, Chopra P. MK2: a novel molecular target for anti-inflammatory therapy. Expert Opin Ther Targets. 2008 Aug;12(8):921-36
Felix RS, Colleoni GW, Caballero OL, Yamamoto M, Almeida MS, Andrade VC, Chauffaille Mde L, Silva WA Jr, Begnami MD, Soares FA, Simpson AJ, Zago MA, Vettore AL. SAGE analysis highlights the importance of p53csv, ddx5, mapkapk2 and ranbp2 to multiple myeloma tumorigenesis. Cancer Lett. 2009 Jun 8;278(1):41-8
Johansen C, Vestergaard C, Kragballe K, Kollias G, Gaestel M, Iversen L. MK2 regulates the early stages of skin tumor promotion. Carcinogenesis. 2009 Dec;30(12):2100-8
Kumar B, Koul S, Petersen J, Khandrika L, Hwa JS, Meacham RB, Wilson S, Koul HK. p38 mitogen-activated protein kinase-driven MAPKAPK2 regulates invasion of bladder cancer by modulation of MMP-2 and MMP-9 activity. Cancer Res. 2010 Jan 15;70(2):832-41
This article should be referenced as such:
Felix R, Alves V, Vettore A, Colleoni G. MAPKAPK2 (mitogen-activated protein kinase-activated protein kinase 2). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):11-14.
Gene Section Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 15
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
MINA (MYC induced nuclear antigen) Makoto Tsuneoka, Kengo Okamoto, Yuji Tanaka
Laboratory of Molecular and Cellular Biology, Department of Molecular Pharmacology, Faculty of
Pharmacy, Takasaki University of Health and Welfare, 60 Nakaorui-machi, Takasaki-shi, Gunma 370-
0033, Japan (MT, KO, YT)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/MINAID44409ch3q11.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI MINAID44409ch3q11.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity Other names: DKFZp762O1912, FLJ14393,
MDIG, MINA53, NO52
HGNC (Hugo): MINA
Location: 3q11.2
Note: MINA (myc induced nuclear antigen) is a
gene whose expression is directly induced by c-
MYC protein. The MINA gene encodes a protein
with a molecular weight of 53 kDa that is localized
in the nucleoplasm and nucleolus.
DNA/RNA
Description
The human MINA gene consists of twelve exons
spanning a 30 kb. The translation start site locates
in exon 2, which follows two distinct exons, exon
1a and exon 1b.
Thus, there are two transcription initiation sites in
the human MINA gene. The exon 1b exists 0.25 kb
downstream of the exon 1a. The stop codon (TAG)
exists in the last exon, exon 10. The open reading
frame of the coding region is 1398 bp, encoding
465 amino acids. mRNA encoding 464 amino acids
(lacking 297Q) is also generated by alternative
splicing due to the lack of the first three bp of exon
7.
Transcription
The human MINA coding sequence consists of
1398 bp from the start codon to the stop codon. In
addition to the 1395 bp-coding sequence, multiple
alternative spliced transcript variants have been
found for this gene. c-MYC protein stimulates the
transcription of MINA through the E-box near the
transcription start sites (Tsuneoka et al., 2002). The
expression of MINA is also induced by serum
(Tsuneoka et al., 2002).
a. Exon-intron structure of the MINA gene. There are two transcription initiation sites at exon 1a and exon 1b. b. mRNA for
human MINA that encodes MINA protein. The protein is coded from exon 2 to exon 10.
MINA (MYC induced nuclear antigen) Tsuneoka M, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 16
Protein
Structural features of MINA protein. The positions of the
JmjC domain is shown (green).
Description
Structure: MINA is a member of the jumonji C
(JmjC) protein family, and suspected to hydroxylate
some proteins to control gene expression.
Activation: The expression of mRNA is elevated
by c-MYC protein, and frequently increased in
various types of cancers, including human colon
cancer, esophageal squamous cell carcinoma
(ESCC). In some types of cancers such as ESCC,
patients with high expression of MINA53 had
shorter survival periods. MINA expression is also
activated not only MYC but also by other factors,
because there are lymphoma and lung cancer tissues
where the expression of MYC is downregulated but
the expression of MINA is elevated (Teye et al.,
2007; Komiya et al., 2010). The expression of
MINA is also activated and mineral dust in human
alveolar macrophage and human lung cancer cell
line, A549 (Zhange et al., 2005).
Expression
MINA is ubiquitously expressed. The expression of
MINA is frequently increased in various types of
human cancers. In mice, MINA expression is high
in some non-neoplastic tissues, including spleen,
thymus, colon and testis, but low in skeletal muscle,
cerebellum, and seminal vesicle. In testis the
expression of MINA is high in spermatogonia and
mitotic prophase cells and weakly in early
pachytene spermatocyte but absent in late
pachytene spermatocytes (Tsuneoka et al., 2006).
Localisation
MINA is diffusely nucleoplasmic and some portion
is accumulated in nucleolus (Tsuneoka et al., 2002).
Function
Specific inhibition of MINA expression suppressed
cell proliferation in some cultured cell lines
(Tsuneoka et al., 2002). Forced expression of
MINA in NIH/3T3 cells induces cell
transformation, and MINA-transfected NIH/3T3
clones produce tumor in nude mice (Komiya et al.,
2010). Therefore, MINA has oncogenic potential.
MINA is a nuclear protein and a member of the
jumonji C (JmjC) protein family. Thus, MINA is
suspected to hydroxylate some proteins to control
gene expression, but its substrate is not clear.
Gene activation: MINA regulates several genes
which are also regulated by MYC. Genes regulated
by MINA but not by MYC include HGF, EGFR,
and IL6 (Komiya et al., 2010).
Gene suppression: Recently, MINA was identified
as a genetic determinant of T(H)2 bias. MINA
specifically binds to and represses the IL4
promoter. MINA overexpression in transgenic mice
impaired IL4 expression, whereas its knockdown in
primary CD4(+) T cells led to IL4 de-repression.
Therefore MINA controls helper T cell
differentiation through an IL4-regulatory pathway
(Okamoto et al,. 2009). These findings suggest that
MINA may play a role on carcinogenesis also in the
field of cancer immunology.
Ribosome biogenesis: MINA is accumulated in
nucleolus (Tsuneoka et al., 2002).
Immunolocalization studies revealed that MINA is
highly concentrated in the granular component of
nucleoli (Eilbracht et al., 2005). MINA is a
constituent of free preribosomal particles but is
absent from cytoplasmic ribosomes. MINA
interacts with various ribosomal proteins as well as
with a distinct set of non-ribosomal nucleolar
proteins. These results suggest that MINA is
directly involved in ribosome biogenesis, most
likely during the assembly process of preribosomal
particles (Eilbracht et al., 2005). MINA was also
suggested to be involved in ribosomal RNA
transcription (Lu et al., 2009).
Homology
The primary sequence of MINA has similarity to
nucleolar protein NO66, which also has a JmjC
domain. The JmjC domain of MINA has 50%
identity to that of NO66. In 2010, it was report that
NO66 directly interacts with Osterix (Osx), which
is an osteoblast-specific transcription factor
required for osteoblast differentiation and bone
formation. NO66 exhibits a JmjC-dependent
histone demethylase activity, which is specific for
both H3K4me and H3K36me in vitro and in vivo. It
was suggested that interactions between NO66 and
Osx regulate Osx-target genes in osteoblasts by
modulating histone methylation states (Sinha et al.,
2010).
Implicated in
Neoplasm diseases
Note
Colon cancer, esophageal squamous cell carcinoma,
gingival squamous cell carcinoma, subtypes of
human lymphoma, renal cell carcinoma,
neuroblastoma, gastric carcinoma, lung cancer and
hepatoma.
Disease
MINA expression is elevated in several types of
carcinomas.
Prognosis
MINA is preferentially expressed in some types of
cancers with a poor prognosis, including
esophageal squamous cell carcinoma, advanced
MINA (MYC induced nuclear antigen) Tsuneoka M, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 17
renal cell carcinoma, neuroblastoma. The
expression levels of MINA may be used as a
prognositic marker in these cancers. On the other
hand, elevated expression of MINA in lung cancer
patients is associated with favorable prognosis.
Colon cancer
Note
The expression of MINA is elevated in all the
adenocarcinomas compared to adjacent non-
neoplastic tissues, which shows little staining.
MINA is expressed in all pathological grades of
cancer as well as in the adenoma. Staining patterns
of Ki-67, a biomarker for cell proliferation, are
similar to those of MINA in most cases. While anti-
Ki-67 antibody strongly stains some well-
proliferating non-neoplastic cells including cells in
the deeper part of the crypts and in lymphoid
germinal centers, antibody to MINA rarely stained
those cells. These results indicate that the elevated
expression of MINA is a characteristic feature in
colon cancer (Teye et al., 2004).
Esophageal cancer
Note
The expression of MINA in tumors is increased
compared with that in adjacent non-neoplastic
tissues. MINA was highly expressed in more than
80% of specimens. Anti-MINA antibody stained
tumors more efficiently than antibody against Ki-
67, a cell proliferation biomarker, in some cancer
specimens. Patients with high expression of MINA
has shorter survival periods, whereas the expression
level of Ki-67 in ESCC shows no relationship to
patient outcome (Tsuneoka et al., 2004).
Primary gingival squamous cell carcinoma
Note
A significant correlation was found between the
expression of MINA and that of Ki-67 in patients
with gingival squamous cell carcinoma or
dysplastic gingiva. No significant correlation was
noted between the expression of MINA or Ki-67
and prognostic factors such as the degree of
differentiation, lymph node metastasis, stage, and
tumor diameter (Kuratomi et al., 2006).
Lymphoma
Note
Although MINA expression is not prominent in
lymphoma in general, it is related to tumor
progression of B cell lymphoma (Teye et al., 2007).
Renal cell carcinoma (RCC)
Note
MINA is expressed in the nuclei of tumor cells and
tubular nuclei of normal renal tissue.
The expression level of MINA is significantly
higher in patients with poor prognostic factors
(stage IV, MVI-positive, and sarcomatoid RCC,
and high Ki-67 LI). The prognosis of high MINA-
expressing tumors was significantly poorer than
that of non-MINA-high tumors (Ishizaki et al.,
2007).
Neuroblastoma
Note
Surgically obtained neuroblastoma specimens were
immunohistochemically stained to determine the
MINA and Cap43 expression levels. A significant
relationship was found between MINA and Ki-67,
between MINA and neurotrophic tyrosine kinase,
receptor, type 1 (TrkA), and between Cap43 and
TrkA. The prognosis is significantly favorable in
the Cap43 high-expression cases, whereas it is
significantly poor in the MINA high-expression
cases (Fukahori et al., 2007).
Gastric carcinomas
Note
Elevated expression of MINA was observed in
91.1% of the gastric carcinomas. No significant
associations were found between MINA and
clinicopathological characteristics such as sex, age,
histological differentiation, distant metastasis and
lymph node metastasis. However, there was a
significant association with depth of invasion and
TMN stage. MINA expression was positively
associated with a proliferation marker, PCNA, level
(Zhang et al., 2008).
Lung cancers
Note
The expression of MINA is elevated in lung cancer
tissues (Lu et al., 2009; Komiya et al., 2010). The
overexpression of MINA is an early event in lung
cancer occurence (Lu et al., 2009; Komiya et al.,
2010). Further, patients with negative staining for
MINA has shorter survival than patients with
positive staining for MINA, especially in stage I or
with squamous cell carcinoma. These results
suggest that overexpression of MINA in lung
cancer patients is associated with favorable
prognosis (Komiya et al., 2009). MINA may inhibit
lung cancer cell invasion (Komiya et al., 2009).
Hepatoma
Note
MINA is diffusely expressed in the nuclei of cancer
cells in the tumor nodule, and is often strong at the
periphery of tumor nodules. MINA expression is
higher in poorly differentiated hepatocellular
carcinoma (HCC) than in well-differentiated HCC,
and there is significant relationship between MINA
expression and histological grade. The high MINA
expression is associated with high expression of a
proliferation marker, antibody to Ki-67. MINA
expression is high in the tumors of > 2 cm of
diameter than in ≤ 2 cm (Ogasawara et al., 2010 in
press).
MINA (MYC induced nuclear antigen) Tsuneoka M, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 18
References Tsuneoka M, Koda Y, Soejima M, Teye K, Kimura H. A novel myc target gene, mina53, that is involved in cell proliferation. J Biol Chem. 2002 Sep 20;277(38):35450-9
Teye K, Tsuneoka M, Arima N, Koda Y, Nakamura Y, Ueta Y, Shirouzu K, Kimura H. Increased expression of a Myc target gene Mina53 in human colon cancer. Am J Pathol. 2004 Jan;164(1):205-16
Tsuneoka M, Fujita H, Arima N, Teye K, Okamura T, Inutsuka H, Koda Y, Shirouzu K, Kimura H. Mina53 as a potential prognostic factor for esophageal squamous cell carcinoma. Clin Cancer Res. 2004 Nov 1;10(21):7347-56
Eilbracht J, Kneissel S, Hofmann A, Schmidt-Zachmann MS. Protein NO52--a constitutive nucleolar component sharing high sequence homologies to protein NO66. Eur J Cell Biol. 2005 Mar;84(2-3):279-94
Zhang Y, Lu Y, Yuan BZ, Castranova V, Shi X, Stauffer JL, Demers LM, Chen F. The Human mineral dust-induced gene, mdig, is a cell growth regulating gene associated with lung cancer. Oncogene. 2005 Jul 21;24(31):4873-82
Kuratomi K, Yano H, Tsuneoka M, Sakamoto K, Kusukawa J, Kojiro M. Immunohistochemical expression of Mina53 and Ki67 proteins in human primary gingival squamous cell carcinoma. Kurume Med J. 2006;53(3-4):71-8
Tsuneoka M, Nishimune Y, Ohta K, Teye K, Tanaka H, Soejima M, Iida H, Inokuchi T, Kimura H, Koda Y. Expression of Mina53, a product of a Myc target gene in mouse testis. Int J Androl. 2006 Apr;29(2):323-30
Fukahori S, Yano H, Tsuneoka M, Tanaka Y, Yagi M, Kuwano M, Tajiri T, Taguchi T, Tsuneyoshi M, Kojiro M. Immunohistochemical expressions of Cap43 and Mina53 proteins in neuroblastoma. J Pediatr Surg. 2007 Nov;42(11):1831-40
Ishizaki H, Yano H, Tsuneoka M, Ogasawara S, Akiba J, Nishida N, Kojiro S, Fukahori S, Moriya F, Matsuoka K,
Kojiro M. Overexpression of the myc target gene Mina53 in advanced renal cell carcinoma. Pathol Int. 2007 Oct;57(10):672-80
Teye K, Arima N, Nakamura Y, Sakamoto K, Sueoka E, Kimura H, Tsuneoka M. Expression of Myc target gene mina53 in subtypes of human lymphoma. Oncol Rep. 2007 Oct;18(4):841-8
Zhang Q, Hu CM, Yuan YS, He CH, Zhao Q, Liu NZ. Expression of Mina53 and its significance in gastric carcinoma. Int J Biol Markers. 2008 Apr-Jun;23(2):83-8
Hemmers S, Mowen KA. T(H)2 bias: Mina tips the balance. Nat Immunol. 2009 Aug;10(8):806-8
Okamoto M, Van Stry M, Chung L, Koyanagi M, Sun X, Suzuki Y, Ohara O, Kitamura H, Hijikata A, Kubo M, Bix M. Mina, an Il4 repressor, controls T helper type 2 bias. Nat Immunol. 2009 Aug;10(8):872-9
Komiya K, Sueoka-Aragane N, Sato A, Hisatomi T, Sakuragi T, Mitsuoka M, Sato T, Hayashi S, Izumi H, Tsuneoka M, Sueoka E. Mina53, a novel c-Myc target gene, is frequently expressed in lung cancers and exerts oncogenic property in NIH/3T3 cells. J Cancer Res Clin Oncol. 2010 Mar;136(3):465-73
Komiya K, Sueoka-Aragane N, Sato A, Hisatomi T, Sakuragi T, Mitsuoka M, Sato T, Hayashi S, Izumi H, Tsuneoka M, Sueoka E. Expression of Mina53, a novel c-Myc target gene, is a favorable prognostic marker in early stage lung cancer. Lung Cancer. 2010 Aug;69(2):232-8
Sinha KM, Yasuda H, Coombes MM, Dent SY, de Crombrugghe B. Regulation of the osteoblast-specific transcription factor Osterix by NO66, a Jumonji family histone demethylase. EMBO J. 2010 Jan 6;29(1):68-79
This article should be referenced as such:
Tsuneoka M, Okamoto K, Tanaka Y. MINA (MYC induced nuclear antigen). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):15-18.
Gene Section Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 19
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
NKX2-1 (NK2 homeobox 1) Theresia Wilbertz, Sebastian Maier, Sven Perner
Institute of Pathology, University Hospital Tubingen, Germany (TW, SM, SP)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/NKX2-1ID44015ch14q13.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI NKX2-1ID44015ch14q13.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity Other names: BCH, BHC, NK-2, NKX2.1,
NKX2A, TEBP, TITF1, TTF-1, TTF1
HGNC (Hugo): NKX2-1
Location: 14q13.3
DNA/RNA
Description
NKX2-1 is regulated by two promoter regions: the
first one is located in intron 1 (5' of exon 1,
regulation of NKX2-1 in lung and thyroid cells).
The second one is situated in the 5' flanking region
of exon 1, it is a 330 bp TATA-less region
containing multiple palindromes and G/C-rich
elements. It regulates NKX2-1 in lung epithelial
cells responding to transcription factors sp1 and
sp3.
Transcription
NKX2-1 is transcribed in two highly conserved
forms: mRNA-isoform 1 contains exon 1, exon 2,
and exon 3, it is translated into a 401 amino acid
protein and represents the minor transcript. mRNA-
isoform 2 is the predominant transcript containing
exon 2 and exon 3. It is translated into a 371 aa
protein.
Figure 1. NKX2-1 gene and NKX2-1 mRNA.
NKX2-1 (NK2 homeobox 1) Wilbertz T, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 20
Figure 2. Upstream and downstream targets of NKX2-1.
Protein
Description
The NKX2-1 protein includes three functional
domains: an N-terminal transactivation domain, a
DNA-binding transactivation domain and a C-
terminal transactivation domain.
Expression
In the lung, expression of NKX2-1 is consistent
throughout all life stages from fetal to adult tissue.
It is expressed in conducting airways type II
alveolar epithelial cells and Clara cells and
uniformly in the terminal respiratory unit.
NKX2-1 expression is also found in thyroid
follicular cells and both normal and hyperplastic C
cells where it activates calcitonin gene expression.
NKX2-1 is not expressed in adult neurons of the
basal ganglia.
During embryonic and fetal development, NKX2-1
expression is found in various tissues (e.g. brain,
lung, thyroid), for details see "function" →
"Embryonic and fetal development".
Localisation
NKX2-1 is a nuclear transcription factor.
Function
In the lung, NKX2-1 regulates the expression of the
lung-specific genes: surfactant protein SP-A, SP-B,
SP-C and Clara cell secretory protein (CCSP).
It cooperates with C/EBPalpha in transactivating
CCSP.
In the transcription of SP-C, NKX2-1 interacts with
nuclear factor I to differentially regulate the
transcription. The longer NKX2-1 isoform reduces
transactivation of SP-C, probably due to some kind
of interference.
NKX2-1 is a key activator of SP-B gene expression
having at least two binding sites at the SP-B
promoter and enhancer. The transactivation
capacity of NKX2-1 regarding the expression of
SP-B is controlled by the sphingolipid ceramide
which is produced in inflammation and reduces
NKX2-1 binding capacity to the SP-B promoter.
SP-B transcription is also inhibited by TGFbeta1-
mediated interaction of smad3 with NKX2-1.
Moreover, NKX2-1 interacts with retinoic acid
receptor (RAR), nuclear receptor coactivators
(p160, CBP/p300) and signal transducers and
activators of transcription 3 (STAT3) in regulation
of SP-B expression.
NKX2-1 (NK2 homeobox 1) Wilbertz T, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 21
Furthermore, NKX2-1 regulates the expression of
the secretoglobulin 3A2 gene (SCGB3A2) in mouse
airways in cooperation with CAATT/enhancer
binding proteins alpha and delta as well as the
expression of ABCA3 which encodes for a lipid
transporter critical for surfactant function at birth
and formation of lamellar bodies.
NKX2-1 also plays an important role in the
endocrine system: it regulates the expression of the
thyroid-specific genes thyroglobulin, thyroid
peroxidase, thyrotropin receptor and sodium-
iodide-symporter, therefore being crucial for proper
thyroid hormone synthesis.
Deletion of NKX2-1 in differentiated neurons of
the hypothalamus in mice causes delayed puberty,
reduced reproductive capacity and a shorter
reproductive span in female mice, suggesting that
NKX2-1 plays an important role in juvenile and
adult endocrine function.
During embryonic and fetal development, NKX2-1
is active in various organs, especially lung, thyroid
and brain.
As a crucial factor for lung development, NKX2-1
is expressed in the ventral foregut endoderm at a
very early stage functioning as a signal which is
essential for specification of a pulmonary cell fate
instead of a liver cell fate. At a later stage, NKX2-1
is critical to the formation of distal pulmonary
structures (whereas proximal lung differentiation is
NKX2-1-independent), a function in which it is
inhibited by TGF-beta.
In addition to that NKX2-1 regulates surfactant
protein genes that are important for the
development of alveolar stability at birth. It induces
SP-A gene expression in fetal lung type II cells
through increased binding of NKX2-1 (mediated by
cAMP) and the NFkappa-B proteins p50 and p65.
Supporting the notion of NKX2-1-dependent SP-
expression, lung and associated respiratory
dysfunction in neonates caused by SP-B-deficiency
are partly induced by down-regulation of NKX2-1.
The main therapeutical option, prenatal
glucocorticoid treatment, induces the expression of
NKX2-1. NKX2-1 regulates expression of
uteroglobin-related protein-1 and claudin-18 during
lung development.
During thyroid gland organogenesis NKX2-1 is
expressed in the ultimobranchial body (UBB) and
in the thyroid diverticulum. It is important for the
survival of UBB-cells and eventually their
dissemination into the thyroid diverticulum and for
the formation of the UBB-derived vesicular
structure. Pendrin and thyroglobulin are
downstream targets of NKX2-1 during thyroid
differentiation. The transactivational activity of
NKX2-1 during thyroid development can be
inhibited by NKX2-5.
In the course of brain development, NKX2-1
expression is found in both telencephalic and
diencephalic domains. It cooperates with Gsh2 to
pattern the ventral telencephalon. Lack of
functional NKX2-1 protein in neurons impairs
developmental differentiation and organization of
basal ganglia and basal forebrain. NKX2-1
upregulates the transcription of nestin, an
intermediate filament protein expressed in
multipotent neuroepithelial cells, by direct binding
to a HRE-CRE-like site (NestBS) within a CNS-
specific enhancer, which indicates that nestin might
be at least one of the effectors of NKX2-1 during
forebrain development.
NKX2-1 expression occurs in neurons of the
arcuate nucleus of the hypothalamus and in glia
cells (tanycytes) in neonatal and adult mice, as well
as in fetal and adult pituicytes suggesting that
NKX2-1 is essential for proper development of the
hypothalamus. Lack of NKX2-1 causes aberrant
trajectory of the dopaminergic pathway in the
developing hypothalamus (mouse-model),
development of GABAergic and cholinergic
neurons is also impaired in NKX2-1 defective mice.
Furthermore, NKX2-1 regulates the specification of
oligodendrocytes and controls the postmitotic
migration of interneurons originating in the medial
ganglionic eminence to either the cortex
(downregulation of NKX2-1) or the striatum
(maintenance of NKX2-1 expression and thus direct
transcriptional activation of neuropilin-2, a
guidance receptor in postmitotic cells). By directly
activating Lhx6 during embryonic development
NKX2-1 plays an essential role for the specification
of cortical interneurons which express parvalbumin
or somatostatin.
In accordance with the findings concerning the role
of NKX2-1 in the development of the above-
mentioned organs, NKX2-1-defective mice die at
birth due to a characteristic set of malformations
and functional impairments: hypoplastic lungs and
insufficient surfactant production, defective
hypothalamus, absence of thyroid and pituitary
gland, delayed development of dopaminergic,
GABAergic and cholinergic neurons.
Mutations
Note
Germinal
Mutations in NKX2-1 (for details see table 1) can
cause benign hereditary chorea (BHC, a dyskinesia,
i.e. a neurological disorder characterized by
abnormal involuntary movements) and brain-lung-
thyroid syndrome (in addition to BHC, patients
suffer from congenital hypothyroidism and infant
respiratory distress syndrome).
A heterozygous substitution at position 1016 in the
coding sequence (C → T) leads to a mutant NKX2-
1 protein (A339V) and can contribute to a
predisposition for multinodular goiter and papillary
thyroid carcinoma.
NKX2-1 (NK2 homeobox 1) Wilbertz T, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 22
Brain-lung-thyroid syndrome congenital hypothyreoidism, infant respiratory distress syndrome, benign hereditary chorea
SNP bp 523 G → T premature stop codon at postition 175
SNP bp 609 C → A premature stop codon at position 145
SNP bp 1320 C → A premature stop codon at position 75
SNP bp 2626 G → T missense mutation: valine → phenylalanine at position 14 of DNA-
binding-domain
SNP splice acceptor
site of intron 2 A → T altered mRNA structure => incorrect removal of introns
Deletion 14q11.2-q13.3
Insertion bp 2595
insertion of GG frameshift mutation: causes truncated protein lacking
the entire third helix of the homeodomain
Cancer predisposition can contribute to predisposition for multinodular goiter and papillary thyroid carcinoma.
SNP bp 1016 C → T missense mutation: A339V
Table 1. Mutations in NKX2-1 gene.
For other heterozygous NKX2-1 mutations in
humans, phenotypes vary widely.
Thyroid dysfunction ranges from mild
hypothyrotrophinaemia to severe congenital
hypothyroidism due to thyroid hypoplasia or even
agenesis. Implication of the lung ranges from a
slight increase in pulmonary infections to severe
neonatal respiratory distress syndrome.
Homozygous NKX2-1 mutations in humans are
probably not viable.
Implicated in
Various cancers
Note
NKX2-1 expression has been found in a variety of
tumor entities, especially lung and thyroid tumors
(for details see table 2).
Lung neoplasms
Disease
NKX2-1 is strongly expressed in 75-90% of
primary lung adenocarcinomas, whereas only 1/4 of
bronchioloalveolar carcinomas show NKX2-1
positivity. Among non-small cell lung cancers,
NKX2-1 is not expressed in squamous cell lung
cancer.
Small cell lung cancer, as well as pulmonary
carcinoids and non-neuroendocrine large-cell
carcinomas partly exhibit NKX2-1 protein
expression.
Prognosis
Overall, NKX2-1 expression is a predictor for
better survival in adenocarcinomas of the lung (just
one smaller study suggested that NKX2-1
expression is associated with poor prognosis).
Controversially, NKX2-1 pathway activation in
lung cancers is associated with poor survival and
cisplatin resistance if PAX9 or Nkx2-8 pathways
are activated at the same time.
Oncogenesis
NKX2-1 is highly amplified in 5-15% of primary
lung adenocarcinomas. In cells harbouring NKX2-1
amplification, this recurrent gene amplification
seems to be a mechanism of high-level NKX2-1
expression.
For a subset of lung adenocarcinomas (especially
those which are derived from the terminal
respiratory unit) sustained expression of NKX2-1
has been shown to be crucial for the survival of
tumor cells. In these tumors RNAi inhibition of
NKX2-1 induces proliferation inhibition, growth
inhibition and apoptosis (lineage-specific
dependency model).
Interestingly, NKX2-1 is also an activator of HOP
(Hsp70/Hsp90 Organizing Protein), a potential
tumor suppressor gene in lung cancer, and it
inhibits EMT (epithelial to mesenchymal
transition). NKX2-1 restores epithelial phenotypes
in lung adenocarcinomas, acting as an adversary of
the EMT-stimulating TGF-beta and a suppressor of
tumor progression and invasiveness. TGF-beta
inhibits the expression of NKX2-1 and thus lung
morphogenesis.
Moreover, NKX2-1 is expressed in most metastatic
lung adenocarcinomas.
Thyroid neoplasms
Disease
Well-differentiated thyroid follicular cell tumors,
such as follicular adenomas, follicular carcinomas
and papillary carcinomas exhibit strong nuclear
positivity for NKX2-1 staining. In contrast,
undifferentiated thyroid carcinomas show low or no
immunoreaction.
NKX2-1 (NK2 homeobox 1) Wilbertz T, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 23
Consistently expressed
Occasionally
expressed Not expressed
Thyroid
- Papillary carcinoma
- Follicular carcinoma
- Medullary carcinoma
- Hurthle cell carcinoma
- Follicular adenoma
- Hyperplastic follicular
cells
- Undifferentiated thyroid
carcinomas
Lung
- Adenocarcinoma
- Small cell lung cancer
(SCLC)
- Pleural effusions of SCLC
- Pulmonary sclerosing
hemangioma
- Bronchioloalveolar
carcinoma (except for
mucinous parts)
- Non-neuroendocrine
large-cell carcinoma
- Signet-ring cell
carcinomas of lung origin
- Pulmonary carcinoids
(50%) - Squamous cell lung cancer
- Pleural mesothelioma
- Bronchioloalveolar
carcinomas (just mucinous
parts)
- Basaloid carcinoma of the
lung
Gastrointestinal system - Small cell cancer of the
esophagus - Colorectal carcinoma
Genitourinary system
- Small cell carcinoma
of the urine bladder
- Nephroblastoma
- Endometrial
carcinoma
- Endocervical
carcinoma
Thymus
- Thymic carcinoma
- Thymoma
Skin
- Merkel cell carcinoma
Neuroectodermal
- Ependymoma
- Glioblastoma - Astrocytoma
- Oligodendroglioma
- Medulloblastoma
- Paraganglioma
- Ganglioglioma
Neuroendocrine
(carcinoid tumorlets,
neuroendocrine cell
hyperplasia, typical
carcinoids, atypical
carcinoids, large cell
neuroendocrine
carcinomas)
- Thyroid origin
- Pulmonary origin - Thymic origin
- Gastrointestinal origin
- Pancreatic origin
- Ovarian origin
- Parathyroid adenoma
- Pituitary adenoma
- Pheochromocytoma
Body cavity fluids
- Lung origin
(adenocarcinoma)
- Genitourinary origin
- Gastrointestinal origin
- Breast origin
- Ovarian origin
Table 2. Expression of NKX2-1 in different tumor entities.
NKX2-1 (NK2 homeobox 1) Wilbertz T, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 24
Concerning parafollicular cells, NKX2-1
expression can be found in normal and hyperplastic
c-cells, as well as in medullary thyroid carcinomas.
However, the signal intensity is much weaker and
less homogenous than observed in tumors
originating from follicular thyroid cells.
Non-malignant branchiogenic cysts can easily be
confounded with papillary thyroid carcinomas.
Since positive immunostaining for NKX2-1 has
been found in a subset of these non-malignant
cervical cysts, NKX2-1 cannot serve to distinguish
between both entities.
Oncogenesis
NKX2-1 is expressed in most differentiated thyroid
neoplasms, but not in undifferentiated tumors of
thyroid origin. On DNA-level, normal thyroids and
papillary carcinomas do not exhibit DNA
methylation in the CpG of NKX2-1 promoter,
whereas undifferenciated thyroid carcinomas show
DNA methylation in this region in about 60%. Most
metastases of thyroid origin are positive for NKX2-
1 expression.
A heterozygous germline mutation, which leads to a
mutant NKX2-1 protein has been shown to be
associated with increased cell proliferation.
Consequently, it might contribute to a
predisposition for multinodular goiter and papillary
thyroid carcinoma (for details see section
mutations).
Neoplasms of the gastrointestinal tract
Disease
Small cell esophageal cancers exhibit NKX2-1
expression in the majority of cases. In contrast,
carcinoids originating from the gastrointestinal
tract, such as ileal, appendical, duodenal,
ampullary, rectal, pancreatic and gastric carcinoids
are negative for NKX2-1 immunohistochemical
staining.
Neoplasms of the genitourinary tract
Disease
NKX2-1 seems to be implicated in neoplasms
arising from the urinary system. Small cell
carcinomas of the urinary bladder are positive for
NKX2-1 staining in 25-40% of cases. Likewise,
large cell neuroendocrine bladder carcinomas
exhibit NKX2-1 expression. In one study, 1/6 of a
set of nephroblastomas showed nuclear positivity
for NKX2-1, whereas metanephric adenomas and
cystic nephromas were NKX2-1 negative.
NKX2-1 expression can be found in benign tubal
and endometrial epithelia, as well as in benign
tumors originating from these tissues. In addition,
malignant tumors of the female genital tract, such
as endocervical adenocarcinomas, small cell
carcinomas of the uterine cervix, endometrioid
adenocarcinomas, serous carcinomas, clear cell
carcinomas, and uterine malignant mixed Mullerian
tumors show positivity for NKX2-1. Staining
morphology in these tumors differs from rare
positive cells to a diffusely positive staining pattern.
Prognosis
No correlation could be detected between positive
NKX2-1 immunostaining in small cell carcinomas
of the urinary bladder and clinicopathologic
features (including outcome, age, sex, smoking
history, stage and metastatic status).
Neuroendocrine neoplasms
Disease
Among well-differentiated neuroendocrine tumors,
only those tumors originating from the lung or
thyroid are positive for NKX2-1 expression.
Neither gastrointestinal typical or atypical
carcinoids, nor neuroendocrine tumors from other
sites (e.g. Merkel cell carcinomas, thymic
carcinoids, ovarian large cell neuroendocrine
carcinomas) show NKX2-1 expression.
Concerning small cell carcinomas, NKX2-1
expression is not specific for small cell lung cancer,
as NKX2-1 expression can also be found in small
cell carcinomas originating from the esophagus,
prostate, bladder or uterine cervix.
Neoplasms of neuroectodermal origin
Disease
NKX2-1 occasionally has been detected in
glioblastoma multiforme and in ependymomas of
the third ventricle. Other primary brain tumors,
such as astrocytomas, oligodendrogliomas,
medulloblastomas and gangliomas from various
sites do not exhibit NKX2-1 expression.
Sellar tumors, including pituicytomas, atypical
pituicytomas, granular cell tumors and spindle cell
oncocytomas can show positive immunostaining for
NKX2-1.
To be noted
Note
NKX2-1 has been well studied in neoplasms of the
lung and thyroid, but lacks a sufficient level of
evidence in other tumor entities.
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Du T, Xu Q, Ocbina PJ, Anderson SA. NKX2.1 specifies cortical interneuron fate by activating Lhx6. Development. 2008 Apr;135(8):1559-67
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Perner S, Wagner PL, Demichelis F, Mehra R, Lafargue CJ, Moss BJ, Arbogast S, Soltermann A, Weder W, Giordano TJ, Beer DG, Rickman DS, Chinnaiyan AM, Moch H, Rubin MA. EML4-ALK fusion lung cancer: a rare acquired event. Neoplasia. 2008 Mar;10(3):298-302
Tomita T, Kido T, Kurotani R, Iemura S, Sterneck E, Natsume T, Vinson C, Kimura S. CAATT/enhancer-binding proteins alpha and delta interact with NKX2-1 to
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synergistically activate mouse secretoglobin 3A2 gene expression. J Biol Chem. 2008 Sep 12;283(37):25617-27
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This article should be referenced as such:
Wilbertz T, Maier S, Perner S. NKX2-1 (NK2 homeobox 1). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):19-28.
Gene Section Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 29
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
NUDT6 (nudix (nucleoside diphosphate linked moiety X)-type motif 6) Leigh-Ann MacFarlane, Paul Murphy
Dalhousie University, Department of Physiology and Biophysics, Faculty of Medicine, 5850 College
Street Sir Charles Tupper Medical Building, Halifax, Nova Scotia B3H 1X5, Canada (LAM, PM)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/NUDT6ID41593ch4q28.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI NUDT6ID41593ch4q28.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity Other names: ASFGF2, bFGF, FGF-2, FGF-AS,
FGF2AS, gfg, gfg-1
HGNC (Hugo): NUDT6
Location: 4q28.1
Note: NUDT6 is a novel nudix protein with
unknown function.
DNA/RNA
Note
Human NUDT6 is located on chromosome 4 in the
region of q28 on the reverse strand, opposite to
FGF-2 gene locus. FGF-2 and NUDT6 genes
overlap at 3' ends, and the mRNAs form a sense-
antisense pair. The NUDT6 mRNA (referred to as
FGF-AS) has been implicated in the regulation of
FGF2 mRNA stability.
Figure A. The schematic representation of the overlap between human NUDT6 (FGF-AS) and FGF2 gene transcripts (colored boxes, coding region; connecting vertical lines, complementary regions between transcripts). Adapted from: MacFarlane LA, et
al., 2010. Molecular Endocrinology 24.
NUDT6 (nudix (nucleoside diphosphate linked moiety X)-type motif 6)
MacFarlane LA, Murphy P
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 30
Figure B. The schematic representation of the human NUDT6 gene transcripts, variants a-g (red boxes, coding region; yellow
boxes, untranslated region).
Description
The human NUDT6 gene is 34271 bp in length,
composed of a 5'UTR, 16 exons, 6 introns and a
3'UTR. The 5' and 3'UTR contain a variety of
regulatory elements that regulate NUDT6
expression. NUDT6 gene transcription is regulated
by a core promoter mapping from -1871 to +181
(relative to the transcription start site +1, up-stream
-), which is 44 kb downstream from the FGF-2
promoter, however the proximal -151/+181 region
confers almost full transcription activity.
The promoter lacks a consensus TATA box or
CCAAT element. The region between the first two
exons contains two Sp1 transcription factor binding
sites (-372/-58 relative to the first exon start site).
The common upstream region (all subsequent
positions relative to first exon start site) from these
start sites contains a multitude of tissue specific
transcription factor binding sites, which include
lymphocyte specific factors Ets at -229 and -83,
GATA at -662 and +56, Lyf-1 at -981; skeletal
muscle consensus E-boxes at -901 and +30; cardiac
factor Nkx-2.5 at -1501 and -582; liver and adipose
C/EBP factor at -624; and testis specific factors
SRY at -1740, -671, +163, +171 and Sox-5 at -
1472, -632.
Two negative regulatory elements also reside in this
shared upstream region, at -1871 and -1315. The
NUDT6 3'UTR contains a singe AU-rich element
(ARE) and seven AU-rich-like sequences which
negatively regulate mRNA stability. A portion of
the NUDT6 coding region and 3'UTR
(+531/+1167) is fully complementary to the 3'UTR
of FGF2 and interaction through this region leads to
the formation of a sense-antisense pair.
Transcription
The primary transcript can be alternatively spliced
to produce at least 7 splice variants, a-d. Full length
variants a and b only differ in the use of an
alternative first exon, designated 1A or 1B. The 3'
ends of the variants share sequence similarity. Two
transcriptional start sites have been identified, one
15 bp upstream of the 1A exon (designated +1) and
the other 84 bp upstream of the 1B exon (+312). It
is unclear whether another transcription start site
specific for other variants are located further
downstream.
NUDT6 transcripts are often designated FGF-AS
(FGF antisense). The two longest transcripts, a and
b, are classified as cis-antisense because they are
transcribed from the same gene locus, on the
opposite DNA strand and their 3'UTR is fully
complementary to the 3'UTR of FGF-2 over two
regions 583 bp and 56 bp in length.
Pseudogene
NA.
Protein
Note
Human NUDT6 encodes 3 novel nudix proteins
with unknown function.
Description
NUDT6 splice variants a-c contain open reading
frames (ORF) that predict isoforms of a novel nudix
motif protein, originally designated GFG. The
nudix box motif is defined by the consensus
signature amino acid sequence
GX5EX7REUXEEXGU,
NUDT6 (nudix (nucleoside diphosphate linked moiety X)-type motif 6)
MacFarlane LA, Murphy P
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 31
Schematic representing predicted NUDT6 isoforms encoded by alternate splice RNA transcripts (deep blue boxes, nudix motif;
light blue box, MTSP-mitochondrial targeting signal peptide).
where X is any amino acid and U is a bulky
hydrophobic amino acid, usually isoleucine, leucine
or valine. To date, three different molecular weight
isoforms have been identified in human, of 35, 28
and 17 kDa, which are presumably generated by
alternative translation initiation. Isoforms are
designated as a, b or c however, this does not
necessarily indicate that the isoform was
synthesized from the corresponding transcript
variant. The 35 kDa isoform is synthesized from the
full length FGF-ASa, by translation initiation from
the in-frame AUG codon located in exon 1A. The
origin of the 28 kDa isoform is unclear. It is
suspected that it is synthesized from an in-frame
CUG codon in exon 2 of either FGF-AS a or b.
However, it is possible that the 28 kDa product is a
proteolytic fragment of the 35 kDa isoform. The 17
kDa isoform may arise from translation initiation at
an in-frame CUG codon in exon 3 of FGF-ASb or
AUG codon in the first exon of FGF-ASc. The
NUDT6 isoforms are detected as stable homo- and
hetero-dimers by western blotting, which can be
disrupted by dithiothreitol (DTT) and boiling.
Potential dimerization domains have been mapped
to both the N-terminus and COOH-terminus of
NUDT6.
Expression
NUDT6 is expressed in a tissue and developmental
stage specific manner. RNA transcripts are detected
in most human tissues including liver, thymus,
spleen, peripheral blood leukocytes, heart skeletal
muscle, testis, colon and kidney. However, which
transcript variants are expressed appears to be tissue
specific. The full length FGF-ASb is thought to be
the predominant variant in most tissues however
variant FGF-ASa is the major variant in normal
hematopoietic tissues. Furthermore, some tissues
co-express FGF-2 and the ratio between FGF-2 and
FGF-AS transcripts varies with tissue and
development stage. FGF-AS levels are relatively
low in many embryonic tissues, with expression
increasing dramatically in a tissue specific manner
postnatally. FGF-2 and FGF-AS exhibit an inverse
relationship in normal tissues, tumor cell lines,
embryonic development and throughout cell cycle
progression.
The level of NUDT6 expression and its ratio with
FGF-2 expression is frequently altered in tumors.
Normal pituitary expresses moderate levels of
NUDT6 and no FGF-2 while pituitary tumors have
reduced NUDT6 expression and high levels of
FGF-2. The NUDT6/FGF-2 expression ratio
decreases dramatically in tumors compared to
normal tissue. Varying NUDT6/FGF-2 ratios have
also been observed in esophageal adenocarcinomas.
Additionally, transient increase in NUDT6
expression occurs in response to treatment with
interleukin-2 and prolactin.
Localisation
NUDT6 can reside in the mitochondria, cytoplasm
and nucleus, however its subcellular localization
varies with isoform, cell type, disease state and
extracellular stimulus. NUDT6a predominantly
localizes to mitochondria whereas NUDT6b and
NUDT6c primarily reside in the cytoplasm and
nucleus. NUDT6 is only found in the cytoplasm of
normal esophageal squamous epithelial cells
whereas in normal lymphocytes it is exclusively
nuclear. However, transformation of these cells
results in the redistribution of NUDT6. Cells from
esophageal adenocarcinoma tumors and lymph
nodes of patient with immunoblastic lymphoma
localize NUDT6 to the nucleus and cytoplasm.
Function
The RNA and protein products appear to have
distinct biological functions. NUDT6 mRNA (FGF-
AS) plays a role in FGF-2 regulation, proliferation,
and cell survival. Additionally, the NUDT6 protein
has been implicated in the control of hormone
production in the pituitary, and possibly in the
removal of potentially hazardous compounds and
metabolites by virtue of its conserved nudix
domain. However, it is not always clear whether a
specific action is a result of the RNA or protein
function and this is further complicated by multiple
antisense splice variants and protein isoforms.
FGF-AS regulates FGF-2 transcript stability.
Although the details of the mechanism involved are
unclear, evidence suggest involvement of RNA
interference and/or a dsRNA duplex formed
between the 3'UTRs of FGF-AS and FGF-2. In
addition to regulating FGF-2 abundance it has been
suggested that it also controls FGF-2 isoform
translation and localization. The regulatory role of
FGF-AS over FGF-2 is thought to account for
observed effects on cell proliferation and survival.
NUDT6 protein is a nudix hydrolase which is a
class of "house cleaning" enzymes capable of
hydrolyzing a broad range of substrates, all defined
as nucleoside diphosphates linked to some other
moiety, that include nucleoside di- and
NUDT6 (nudix (nucleoside diphosphate linked moiety X)-type motif 6)
MacFarlane LA, Murphy P
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 32
triphosphates, dinucleoside and diphosphoinositol
polyphosphates, nucleotide sugars and RNA caps.
The substrate of human NUDT6 has yet to be
elucidated and therefore its physiological function
remains unknown. NUDT6 has observed effects on
cell proliferation independent of those associated
with FGF-AS. NUDT6 overexpression in human
colorectal cancer cells increases proliferation.
Perhaps NUDT6's effects on proliferation are
dependent on expression level, isoform and/or cell
type, as is the case with FGF-2. Furthermore,
NUDT6 is involved in hormone production. GFG
expression can increase levels of prolactin.
However it is unclear if these effects are mediated
through the same MAPK pathway utilized by FGF-
2 to increase prolactin expression.
Additionally, NUDT6 alters the isoform ratio of
growth hormone, by increasing synthesis of the 22
kDa isoform and not the 20 kDa.
Homology
NUDT6 contains a conserved nudix motif common
to other members of the Nudix family of
phosphohydrolases. The Nudix motif is
GXXXXXEXXXXXXXREUXEEXGU where U is
Isoleucine, Leucine, or Valine and X is any amino
acid. NUDT6 is highly conserved among man, cow,
mouse, worm, and fruit fly, and GFG homologs
across species are more evolutionarily related to
each other than to other nudix proteins from the
same species.
Implicated in
Esophageal adenocarcinoma
Note
Esophageal adenocarcinoma refers to uncontrolled
growth of glandular cells in the esophagus and the
junction between the esophagus and the stomach.
Prognosis
Elevated expression of FGF-AS in FGF-2
expressing esophageal adenocarcinomas is
associated with reduced tumor reoccurrence
following surgical resection of tumors and
increased survival rates, suggesting that it may be
used as a prognostic indicator.
Oncogenesis
Esophageal adendocarcinoma tumors overexpressed
FGF-AS and cytoplasmic GFG in comparison to
normal match esophageal tissue. However, the
reduced tumor reoccurrence and improved survival
rates specifically correlated to FGF-AS levels, not
GFG levels. Evidence suggests FGF-AS tumor
suppressive role is a result of its post-transcription
control over FGF-2.
Melanoma
Note
Melanoma is a malignant tumor of melanocytes,
which are found primarily in the skin, however they
can develop in melanocytes found in the eye and
bowel. A characteristic of aggressive melanomas is
their ability to form fluid-conducting vasculogenic-
like networks.
Oncogenesis
A preliminary study investigating these 3D tubular
networks within tumors identified NUDT6 as one
of the many genes overexpressed in aggressive
melanomas and speculated it is involved in
promoting self-renewal and tumor cell plasticity in
melanoma cancer networks. Additionally, they
suggest FGF-AS could play a role in the
development of the endothelia-lined vasculature
networks in melanomas indirectly through its
regulatory control over FGF-2 expression, which is
associated with angiogenesis, proliferation and
survival.
Colorectal cancer
Note
Colorectal cancer refers to uncontrolled growth of
cells that line the colon, rectum and appendix,
collectively the large intestine.
Oncogenesis
Induced overexpression of NUDT6 in a variety of
human colorectal cells significantly increases
cancer cell proliferation and their clonogenic
capacity. NUDT6 is described as having tumor
promoting functions in this cellular environment
and it is suggested that it plays a role in colorectal
cancer development and progression.
Endometriosis
Note
Endometriosis is a medical condition affecting the
endometrium lining of the uterus. The endometrium
is comprised of hormonally responsive cells that
proliferate and secrete under the influence of
estrogen and progesterone. Upon menstruation the
endometrium lining is shed as a part of the
menstrual flow. Endometriosis describes the
presence of endometrial cells outside of the uterus,
such as the ovaries, fallopian tubes, bladder and
interstitial space in the abdominal cavity.
Patients with endometriosis lesions have reduced
FGF-AS-b mRNA levels and elevated FGF-2
mRNA levels during the late proliferative phase of
the menstrual cycle, compared to control patients.
This increased FGF/FGF-AS ratio is thought to
contribute to the development of endometriosis.
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Murphy PR, Knee RS. Identification and characterization of an antisense RNA transcript (gfg) from the human basic fibroblast growth factor gene. Mol Endocrinol. 1994 Jul;8(7):852-9
NUDT6 (nudix (nucleoside diphosphate linked moiety X)-type motif 6)
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Bessman MJ, Frick DN, O'Handley SF. The MutT proteins or "Nudix" hydrolases, a family of versatile, widely distributed, "housecleaning" enzymes. J Biol Chem. 1996 Oct 11;271(41):25059-62
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Baguma-Nibasheka M, Li AW, Osman MS, Geldenhuys L, Casson AG, Too CK, Murphy PR. Coexpression and regulation of the FGF-2 and FGF antisense genes in leukemic cells. Leuk Res. 2005 Apr;29(4):423-33
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factor for esophageal cancer recurrence and reduced survival, which is ameliorated by coexpression of the FGF-2 antisense gene. Clin Cancer Res. 2005 Nov 1;11(21):7683-91
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This article should be referenced as such:
MacFarlane LA, Murphy P. NUDT6 (nudix (nucleoside diphosphate linked moiety X)-type motif 6). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):29-33.
Gene Section Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 34
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
PARVB (parvin, beta) Cameron N Johnstone
Cancer Metastasis Laboratory, Research Division, Peter MacCallum Cancer Centre, 2 St Andrew's
Place, East Melbourne, 3002, Victoria, Australia (CNJ)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/PARVBID46486ch22q13.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI PARVBID46486ch22q13.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity
Other names: CGI-56, affixin, beta-parvin
HGNC (Hugo): PARVB
Location: 22q13.31
Local order: PARVB is located telomeric to the
SAMM50 gene and centromeric to the PARVG
gene at 22q13.31.
DNA/RNA
Note
Genethon marker D22S1171 is located at the 5' end
of the gene (Mongroo et al., 2004). Genethon
marker D22S1171 is located between exon 2 and
exon 1A of the PARVB gene.
The PARVA gene is located at 11p15.3.
Figure A. Generation of transcript diversity by alternative promoter usage. Horizontal lines above the gene structure indicate
human genomic DNA BAC clones. The NCBI accession numbers of the clones, and clone names (in brackets) are shown. Figure adapted from Mongroo et al., 2004.
PARVB (parvin, beta) Johnstone CN
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 35
Figure B. Human polyA
+ RNA Multiple Tissue Northern blot (Origene) probed with full-length PARVB1 cDNA probe radiolabeled
to a specific activity of > 5 x 108 cpm / mg (Johnstone C.N., unpublished). The two PARVB mRNA transcripts are indicated. The
higher M.W. band most likely corresponds to non-specific hybridization (n/s).
Description
PARVB3/CLINT, which encodes the longer
Parvin-beta protein isoform is transcribed from
promoter 1 and contains two additional 5' exons
(exons 1 and 2) not present in PARVB1, and
comprises 14 exons in total. PARVB1 encodes the
shorter Parvin-beta protein isoform, is transcribed
from promoter 1A, and comprises 13 exons in total.
Both promoters contain CpG islands that span the
transcription start sites. PARVB3/CLINT contains
70 unique N-terminal amino acids not present in the
short isoform. (See figure A).
Transcription
As with PARVA, human PARVB mRNA
expression is highest in heart, followed by skeletal
muscle, where it localises to the sarcolemma
(Yamaji et al., 2001; Matsuda et al., 2005). Both
PARVB mRNA transcripts are essentially
ubiquitously expressed (Korenbaum et al., 2001),
but with lower expression in gastrointestinal tissues
(stomach, small intestine, colon). (See figure B).
Protein
Description
The major functional domains of Parvin-beta are
two 'atypical' calponin homology (CH) domains,
termed CH1 (106 amino acids) and CH2 (107
amino acids). Each CH domain contains two actin
binding sequences (ABS), although Parvin-beta has
not been shown to bind actin directly (Korenbaum
et al., 2001; Sepulveda and Wu, 2006). Parvin-beta
physically interacts with Dysferlin and ARHGEF6
(alpha-PIX) through the CH1 domain (Matsuda et
al., 2005; Rosenberger et al., 2003) and with ILK
and alpha-actinin through the CH2 domain (Yamaji
et al., 2001; Yamaji et al., 2004). Parvin-beta was
also recently reported to directly interact with AKT
(Kimura et al., 2010).
Expression
PARVB is essentially ubiquitously expressed.
Localisation
Parvin-beta localises to focal adhesions but also to
the nucleus, which is most likely due to NLS motifs
in the N-terminal region (Mongroo et al., 2004;
Johnstone et al., 2008). Parvin-beta is incorporated
into focal adhesions as part of the heterotrimeric
'IPP complex'. The ternary complex contains 1
molecule of integrin linked kinase (ILK), 1 Parvin
isoform, and 1 PINCH (LIMS) isoform, (Legate et
al., 2006). Binding of Parvin-alpha and Parvin-beta
to the kinase domain of ILK is mutually exclusive
(Zhang et al., 2004). Formation of the IPP complex
also dictates total protein levels of each component,
as any excess ILK, Parvin, or PINCH not
incorporated into IPP is degraded in a proteasome-
dependent manner (Fukuda et al., 2003).
Depiction of functional domains of Parvin-beta(long) and Parvin-beta(short). NLS, nuclear localization sequence; ABS,
actin binding sequence; CH, calponin homology. Adapted from Sepulveda and Wu, 2006.
PARVB (parvin, beta) Johnstone CN
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 36
Function
Parvin-beta participates in focal adhesion dynamics
through involvement in the IPP complex. The high
expression levels in cardiac and skeletal muscle
suggest important function(s) in these organs. In
skeletal muscle, it binds dysferlin at the
sarcolemma and thus may be involved with
membrane repair (Yamaji et al., 2001; Matsuda et
al., 2005; Legate et al., 2006). Parvin-beta and
Parvin-alpha appear to negatively regulate the
expression of each other (Zhang et al., 2004;
Johnstone et al., 2008). Parvin-beta may modulate
signalling through ILK as overexpression of Parvin-
beta reduced AKT (S473) and GSK3beta (S9)
phosphorylation in response to EGF stimulation
(Mongroo et al., 2004). Parvin-beta was recently
reported to directly interact with AKT (Kimura et
al., 2010), which may explain its effects on AKT
phosphorylation. Parvin-beta interacts with
ARHGEF6 (alpha-PIX), an exchange factor for
RAC1, thus implicating Parvin-beta in regulation of
RAC signalling downstream of integrin
engagement (Rosenberger et al., 2003). Finally,
Parvin-beta may affect metabolic pathways through
promotion of CDK9-mediated phosphorylation and
activation of PPARgamma transcriptional activity
in the nucleus (Johnstone et al., 2008).
Interestingly, Parvb knockout mice were recently
generated. Whilst constitutive Parva null mice
feature kidney and cardiovascular defects and die
between E10.5 and E14.5 (Lange et al., 2009;
Montanez et al., 2009), constitutive Parvb null mice
are viable (Wickström et al., 2010), although a
detailed phenotypic analysis has not yet been
described.
Homology
Human Parvin-beta is most closely related to
Parvin-alpha [75% identity with Parvin-beta(short)
and 67% identity with Parvin-beta(long)] and more
distantly to Parvin-gamma [41% identity with both
Parvin-beta(short) and Parvin-beta(long)].
Mutations
Note
No mutations reported to date.
Germinal
Germline SNPs are identified in the PARVB gene
by direct sequencing of PCR products amplified
from cDNA prepared from 16 primary ductal
adenocarcinomasand adjacent normal mammary
gland from the same patient. Two non-synonymous
SNPs were identified, W37R, and E175K
(Johnstone et al., manuscript in preparation).
Somatic
No somatic mutations were found in an analysis of
16 breast adenocarcinomas as presented above
(Johnstone et al., manuscript in preparation).
According to the C.O.S.M.I.C. online database
(Forbes et al., 2008), 171 unique cancer samples
have been analysed for alterations in the PARVB
gene, with no somatic changes found. A breakdown
of the samples analysed is given below.
Name Alleles Location Base Position† Amino Acid
Position‡
Amino Acid
change
No. of
Alleles
A98C A/C Intron 1 98** n/a n/a 3/8
W37R T/C Exon 2 252 37 W>R 3/8
D150D C/T Exon 5 593 150 D>D 2/32
E175K G/A Exon 6 666 175 E>K 2/32
A223A C/T Exon 7 812 223 A>A 2/32
G316G^ C/T Exon 12 1097 318 G>G 2/32
F354F^ C/T Exon 13 1205 354 F>F 2/32
† Relative to transcription start site
‡ Relative to translation start site
^ Occur as a haplotype
** Relative to splice site
Cancer Type No. of Specimens Reference
Breast 11 Sjöblom et al., 2006
Glioma 23 Parsons et al., 2008
Clear Cell Renal 101 Dalgliesh et al., 2010
Colon 12 Sjöblom et al., 2006
Lung (cell lines) 11 N/A
PARVB (parvin, beta) Johnstone CN
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 37
Pancreas (cell lines) 1 N/A
Mesothelioma (cell lines) 1 N/A
Melanoma (cell lines) 6 N/A
Urinary tract (cell lines) 2 N/A
HNSCC (cell lines) 3 N/A
Implicated in
Breast cancer
Note
Parvin-beta mRNA levels are reduced in primary
human ductal adenocarcinoma compared with
adjacent normal mammary gland. PARVB mRNA
levels are also reduced in MDA-MB-231 and
MDA-MB-453 cell lines. Post-transcriptional
downregulation of protein expression may also
occur in cancer cells such as MCF7 (Mongroo et
al., 2004). Ectopic Parvin-beta expression in MDA-
MB-231 metastatic breast cancer cells increased
adhesion and reduced invasion. Ectopic expression
also reduced tumorigenicity of the same cell line in
nude mice in vivo. Parvin-beta expression did not
affect proliferation of the cells in vitro, but reduced
Ki-67 staining was observed in Parvin-beta
transfectants in vivo (Johnstone et al., 2008).
Parvin-beta overexpression was also reported to
promote apoptosis in HeLa cervical cancer cells
(Zhang et al., 2004).
Prognosis
Association with prognosis has not been studied to
date.
References Korenbaum E, Olski TM, Noegel AA. Genomic organization and expression profile of the parvin family of focal adhesion proteins in mice and humans. Gene. 2001 Nov 14;279(1):69-79
Yamaji S, Suzuki A, Sugiyama Y, Koide Y, Yoshida M, Kanamori H, Mohri H, Ohno S, Ishigatsubo Y. A novel integrin-linked kinase-binding protein, affixin, is involved in the early stage of cell-substrate interaction. J Cell Biol. 2001 Jun 11;153(6):1251-64
Fukuda T, Chen K, Shi X, Wu C. PINCH-1 is an obligate partner of integrin-linked kinase (ILK) functioning in cell shape modulation, motility, and survival. J Biol Chem. 2003 Dec 19;278(51):51324-33
Rosenberger G, Jantke I, Gal A, Kutsche K. Interaction of alphaPIX (ARHGEF6) with beta-parvin (PARVB) suggests an involvement of alphaPIX in integrin-mediated signaling. Hum Mol Genet. 2003 Jan 15;12(2):155-67
Mongroo PS, Johnstone CN, Naruszewicz I, Leung-Hagesteijn C, Sung RK, Carnio L, Rustgi AK, Hannigan GE. Beta-parvin inhibits integrin-linked kinase signaling and is downregulated in breast cancer. Oncogene. 2004 Nov 25;23(55):8959-70
Yamaji S, Suzuki A, Kanamori H, Mishima W, Yoshimi R, et al. Affixin interacts with alpha-actinin and mediates integrin signaling for reorganization of F-actin induced by
initial cell-substrate interaction. J Cell Biol. 2004 May 24;165(4):539-51
Zhang Y, Chen K, Tu Y, Wu C. Distinct roles of two structurally closely related focal adhesion proteins, alpha-parvins and beta-parvins, in regulation of cell morphology and survival. J Biol Chem. 2004 Oct 1;279(40):41695-705
Matsuda C, Kameyama K, Tagawa K, Ogawa M, Suzuki A, et al. Dysferlin interacts with affixin (beta-parvin) at the sarcolemma. J Neuropathol Exp Neurol. 2005 Apr;64(4):334-40
Legate KR, Montañez E, Kudlacek O, Fässler R. ILK, PINCH and parvin: the tIPP of integrin signalling. Nat Rev Mol Cell Biol. 2006 Jan;7(1):20-31
Sepulveda JL, Wu C. The parvins. Cell Mol Life Sci. 2006 Jan;63(1):25-35
Sjöblom T, Jones S, Wood LD, Parsons DW, Lin J, et al. The consensus coding sequences of human breast and colorectal cancers. Science. 2006 Oct 13;314(5797):268-74
Forbes SA, Bhamra G, Bamford S, Dawson E, Kok C, Clements J, Menzies A, Teague JW, Futreal PA, Stratton MR. The Catalogue of Somatic Mutations in Cancer (COSMIC). Curr Protoc Hum Genet. 2008 Apr;Chapter 10:Unit 10.11
Johnstone CN, Mongroo PS, Rich AS, Schupp M, et al. Parvin-beta inhibits breast cancer tumorigenicity and promotes CDK9-mediated peroxisome proliferator-activated receptor gamma 1 phosphorylation. Mol Cell Biol. 2008 Jan;28(2):687-704
Parsons DW, Jones S, Zhang X, Lin JC, Leary RJ, et al. An integrated genomic analysis of human glioblastoma multiforme. Science. 2008 Sep 26;321(5897):1807-12
Lange A, Wickström SA, Jakobson M, Zent R, Sainio K, Fässler R. Integrin-linked kinase is an adaptor with essential functions during mouse development. Nature. 2009 Oct 15;461(7266):1002-6
Montanez E, Wickström SA, Altstätter J, Chu H, Fässler R. Alpha-parvin controls vascular mural cell recruitment to vessel wall by regulating RhoA/ROCK signalling. EMBO J. 2009 Oct 21;28(20):3132-44
Dalgliesh GL, Furge K, Greenman C, Chen L, Bignell G, et al. Systematic sequencing of renal carcinoma reveals inactivation of histone modifying genes. Nature. 2010 Jan 21;463(7279):360-3
Kimura M, Murakami T, Kizaka-Kondoh S, Itoh M, et al. Functional molecular imaging of ILK-mediated Akt/PKB signaling cascades and the associated role of beta-parvin. J Cell Sci. 2010 Mar 1;123(Pt 5):747-55
Wickström SA, Lange A, Montanez E, Fässler R. The ILK/PINCH/parvin complex: the kinase is dead, long live the pseudokinase! EMBO J. 2010 Jan 20;29(2):281-91
This article should be referenced as such:
Johnstone CN. PARVB (parvin, beta). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):34-37.
Gene Section Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 38
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
PIAS3 (protein inhibitor of activated STAT, 3) Gilles Spoden, Werner Zwerschke
Institute for Medical Microbiology and Hygiene, University Medical Center of the Johannes
Gutenberg University Mainz, Hochhaus am Augustus-platz, 55131 Mainz, Germany (GS); Cell
Metabolism and Differentiation Research Group, Institute for Biomedical Aging Research, Rennweg
10, 6020 Innsbruck, Austria (ZW); Tumorvirology Research Group, Tyrolean Cancer Research
Institute at Medical University Innsbruck, Innrain 66, 6020 Innsbruck, Austria (ZW)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/PIAS3ID41709ch1q21.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI PIAS3ID41709ch1q21.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity
Other names: FLJ14651, KChAP, ZMIZ5
HGNC (Hugo): PIAS3
Location: 1q21.1
DNA/RNA
Note
The gene codes for a protein of the PIAS family
(protein inhibitor of activated STAT (signal
transducer and activator of transcription)). PIAS3
regulates the activity of several transcription factors
by direct protein-protein interaction. Further,
PIAS3 is a SUMO (small ubiquitin-like modifier)-
E3 ligase, catalyzing the covalent, post-translational
modification of specific target proteins with
SUMO. Different splice variants of PIAS3 have
been identified but the full-length sequence of some
of these variants has not been described.
Description
The human PIAS3 gene is 10559 bp long and
consists of 14 exons and 13 introns.
Transcription
Transcript length 2902 bp (CDS 1887 bp ; residues
628 aa).
Pseudogene
No pseudogene reported.
Figure 1. PIAS3 gene 10559 bp. Exons 1 to 14 (UTR in white, coding sequence in red) Exon 1: 1-115 (5'UTR: 1-91); Exon 2:
2075-2492; Exon 3: 2650-2734; Exon 4: 2963-3013; Exon 5: 3255-3345; Exon 6: 4201-4335; Exon 7: 4518-4623; Exon 8: 5195-5268; Exon 9: 5417-5577; Exon 10: 7928-8061; Exon 11: 8142-8310; Exon 12: 8495-8628; Exon 13: 8812-8849; Exon 14: 9369-
10559 (3'UTR: 9636-10559).
Figure 2. The schematic domain structure of human PIAS3 protein is shown. SAP domain: nuclear localization and binding to
DNA, transcription factors, coregulators. PINIT: nuclear retention, transcriptional repression. SP-RING: protein-protein interactions, interacts with the SUMO conjugase Ubc9, sumoylation. SIM: binding to SUMO. S/T: variable region, binding to
coactivators.
PIAS3 (protein inhibitor of activated STAT, 3) Spoden G, Zwerschke W
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 39
Protein
Description
The human PIAS3 protein is a E3 SUMO-protein
ligase consisting of 628 amino acids. It contains 5
conserved regions, the SAP, PINIT, SP-RING, SIM
and S/T domains (figure 2).
Expression
PIAS3 is ubiquitously expressed.
Localisation
Nuclear as well as cytoplasmic localization.
Function
PIAS3 belongs to the mammalian protein inhibitor
of activated STAT (PIAS) protein family, originally
identified as cytokine-induced inhibitors of the
STAT family of transcription factors. This protein
class, referred to as SUMO-E3 ligases, increases the
efficiency of SUMO conjugation. SUMO, a small
ubiquitin-like modifier protein, is conjugated to a
large number of cellular target proteins. Similar to
enzymatic ubiquitination, the conjugation of
specific SUMO proteins (SUMO-1-SUMO-3) to
target proteins requires an E1-activating enzyme
(Aos1/Uba2) as well as an E2-type SUMO-1-
conjugating enzyme (Ubc9). Similar to many
ubiquitin E3 ligases, these proteins contain a
putative RING finger-like structure (SP-RING,
figure 2), which is essential for their SUMO-E3
ligase activities toward various target proteins.
Sumoylation is a dynamic process with highly
diverse outcomes, ranging from changes in
subcellular localization, signal transduction,
transcriptional regulation to altered activity and
stability of the modified protein. PIAS3 do,
however, not only operate as SUMO-E3, since its
coregulator effects are often independent of its
RING-finger like domain but dependent on its
capability to interact with sumoylated proteins via
its conserved SIM (SUMO-interacting motif) or
SAP (scaffold attachment factor-A/B/acinus/PIAS)
domain (figure 2). Beside the N-terminal SAP, the
SIM and the RING-type zinc-binding domain, a
PINIT motif, and a serine/ threonine-rich C-
terminal region (S/T) is conserved in PIAS3 (figure
2).
PIAS3 is involved in cytoplasmic regulation, such
as functional interaction of PIAS3 with
metabotropic glutamate receptor-8, voltage-gated
potassium channel Kv1.5 and pyruvate kinase
subtype M2, but the majority of so far reported
interactions of the PIAS3 protein occurred with
transcription factors or other proteins linked to
nuclear regulation. PIAS3 can act in both
transcriptional repression and activation. PIAS3 has
been shown to repress STAT3 and Stat5 dependent
transcriptional activation by blocking the DNA-
binding of the factor without influencing its
sumoylation. It interacts with and promotes
sumoylation of the photoreceptor-specific
transcription factor Nr2e3 when bound to specific
promoters, which converts the factor to a
transcriptional repressor. Moreover, PIAS3 was
described as a repressor of microphthalmia
transcription factor (MITF) and it was shown that
PIAS3 blocks NF-kB mediated transcriptional
activation by interacting with the p65/RelA subunit.
Repression of IRF1-mediated transcription by
PIAS3 has also been shown. PIAS3 has been shown
to activate transcription mediated by Smad proteins
through forming a complex with Smads and
coactivator p300/CBP; moreover, PIAS proteins
enhance steroid receptor-dependent transcription
through an SP-RING-mediated interaction and
sumoylation of the coactivator protein
GRIP1/SCR2. Finally, PIAS3 was shown to
modulate the ability of TIF2 to mediate ligand-
enhanced transcription activation positively or
negatively, for different steroid receptors.
Homology
The mammalian PIAS family consists of seven
structurally related proteins (PIAS1, PIAS3,
PIAS3b, PIASxa, PIASxb, PIASy, and PIASyE6)
encoded by four genes. PIAS orthologs are found in
nonvertebrate animal species, plants and yeasts.
Implicated in
Prostate Cancer
Oncogenesis
PIAS3 is expressed in normal prostate and in
prostate cancer cells and has been shown to
modulate the transcriptional activity of androgen
receptor in prostate cancer cells. Moreover, PIAS3
(KChAP) induces increased K+ efflux and
apoptosis in prostate cancer lines.
Glioblastoma multiforme (GBM)
Oncogenesis
The activation of STATs and loss of their natural
inhibitors SOCS and PIAS is common in various
human cancers. STAT3, a cytoplasmic transcription
factor that becomes activated in response to a
variety of cytokines and growth factors is aberrantly
activated in GBM tumors. STAT3 activation
correlates with strongly reduced PIAS3 protein
expression in GBM tissues. Inhibition of PIAS3
resulted in enhanced glioblastoma cellular
proliferation, and, conversely, PIAS3
overexpression inhibits STAT3 transcriptional
activity, expression of STAT3-regulated genes, and
cell proliferation. This suggests that the loss of
PIAS3 in GBM contributes to enhanced STAT3
transcriptional activity and subsequent cell
proliferation.
PIAS3 (protein inhibitor of activated STAT, 3) Spoden G, Zwerschke W
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 40
Melanoma
Oncogenesis
PIAS3 functions as a key molecule in suppressing
the transcriptional activity of both MITF and
STAT3, two transcription factors that play a major
role in the development, proliferation and survival
of mast cells and melanocytes. In addition to its role
in normal cell signaling, constitutively activated
STAT3 signaling directly contributes to
oncogenesis in many human cancers. STAT3
cooperates with MITF in the induction of cellular
transformation. Evidence was provided suggesting
that PIAS3 halt proliferation and induce apoptotic
cell death in mast cells and in melanoma cells by
inhibiting the transcriptional activity of the two
oncogenic factors MITF and STAT3. Therefore
PIAS3 may play a role in tumor suppression by
inhibiting oncogenic processes induced by STAT3
and MITF.
Non-small cell lung cancer (NSCLC)
Disease
The Epidermal Growth Factor Receptor (EGFR)-
STAT3 axis plays an important role in oncogenic
signaling of non-small cell lung cancer (NSCLC).
The negative regulator of STAT3-mediated
transcriptional activation, PIAS3, was shown to
modulate oncogenic EGFR-STAT3 signaling in
lung cancer. Overexpression of PIAS3 decreases
STAT3 transcriptional activity and proliferation of
NSCLC cells and when used in conjunction with
EGFR inhibitors, further increased the anti-
proliferative effects. This suggests that PIAS3 acts
as an inhibitor of EGFR-STAT3 induced oncogenic
action.
To be noted
Note
TAR syndrome (Thrombocytopenia-absent radius)
is a rare genetic disorder characterized by low
platelet counts and bilateral radial aplasia. TAR is
also frequently associated with cardiac
abnormalities and cow's milk intolerance. In 2007 a
research article described a common microdeletion
of 200 kb on chromosome 1q21.1 in patients with
TAR syndrome. PIAS3 is one of 11 genes
encompassed by this microdeletion.
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Wible BA, Yang Q, Kuryshev YA, Accili EA, Brown AM. Cloning and expression of a novel K+ channel regulatory protein, KChAP. J Biol Chem. 1998 May 8;273(19):11745-51
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Minty A, Dumont X, Kaghad M, Caput D. Covalent modification of p73alpha by SUMO-1. Two-hybrid screening with p73 identifies novel SUMO-1-interacting proteins and a SUMO-1 interaction motif. J Biol Chem. 2000 Nov 17;275(46):36316-23
Gross M, Liu B, Tan J, French FS, Carey M, Shuai K. Distinct effects of PIAS proteins on androgen-mediated gene activation in prostate cancer cells. Oncogene. 2001 Jun 28;20(29):3880-7
Hari KL, Cook KR, Karpen GH. The Drosophila Su(var)2-10 locus regulates chromosome structure and function and encodes a member of the PIAS protein family. Genes Dev. 2001 Jun 1;15(11):1334-48
Hochstrasser M. SP-RING for SUMO: new functions bloom for a ubiquitin-like protein. Cell. 2001 Oct 5;107(1):5-8
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Kotaja N, Vihinen M, Palvimo JJ, Jänne OA. Androgen receptor-interacting protein 3 and other PIAS proteins cooperate with glucocorticoid receptor-interacting protein 1 in steroid receptor-dependent signaling. J Biol Chem. 2002 May 17;277(20):17781-8
Levy C, Nechushtan H, Razin E. A new role for the STAT3 inhibitor, PIAS3: a repressor of microphthalmia transcription factor. J Biol Chem. 2002 Jan 18;277(3):1962-6
Nakagawa K, Yokosawa H. PIAS3 induces SUMO-1 modification and transcriptional repression of IRF-1. FEBS Lett. 2002 Oct 23;530(1-3):204-8
Nishida T, Yasuda H. PIAS1 and PIASxalpha function as SUMO-E3 ligases toward androgen receptor and repress androgen receptor-dependent transcription. J Biol Chem. 2002 Nov 1;277(44):41311-7
Rycyzyn MA, Clevenger CV. The intranuclear prolactin/cyclophilin B complex as a transcriptional inducer. Proc Natl Acad Sci U S A. 2002 May 14;99(10):6790-5
Tan JA, Hall SH, Hamil KG, Grossman G, Petrusz P, French FS. Protein inhibitors of activated STAT resemble scaffold attachment factors and function as interacting nuclear receptor coregulators. J Biol Chem. 2002 May 10;277(19):16993-7001
Wible BA, Wang L, Kuryshev YA, Basu A, Haldar S, Brown AM. Increased K+ efflux and apoptosis induced by the potassium channel modulatory protein KChAP/PIAS3beta in prostate cancer cells. J Biol Chem. 2002 May 17;277(20):17852-62
Duval D, Duval G, Kedinger C, Poch O, Boeuf H. The 'PINIT' motif, of a newly identified conserved domain of the PIAS protein family, is essential for nuclear retention of PIAS3L. FEBS Lett. 2003 Nov 6;554(1-2):111-8
Levy C, Sonnenblick A, Razin E. Role played by microphthalmia transcription factor phosphorylation and its
PIAS3 (protein inhibitor of activated STAT, 3) Spoden G, Zwerschke W
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 41
Zip domain in its transcriptional inhibition by PIAS3. Mol Cell Biol. 2003 Dec;23(24):9073-80
Schmidt D, Müller S. PIAS/SUMO: new partners in transcriptional regulation. Cell Mol Life Sci. 2003 Dec;60(12):2561-74
Jang HD, Yoon K, Shin YJ, Kim J, Lee SY. PIAS3 suppresses NF-kappaB-mediated transcription by interacting with the p65/RelA subunit. J Biol Chem. 2004 Jun 4;279(23):24873-80
Johnson ES. Protein modification by SUMO. Annu Rev Biochem. 2004;73:355-82
Joo A, Aburatani H, Morii E, Iba H, Yoshimura A. STAT3 and MITF cooperatively induce cellular transformation through upregulation of c-fos expression. Oncogene. 2004 Jan 22;23(3):726-34
Long J, Wang G, Matsuura I, He D, Liu F. Activation of Smad transcriptional activity by protein inhibitor of activated STAT3 (PIAS3). Proc Natl Acad Sci U S A. 2004 Jan 6;101(1):99-104
Sonnenblick A, Levy C, Razin E. Interplay between MITF, PIAS3, and STAT3 in mast cells and melanocytes. Mol Cell Biol. 2004 Dec;24(24):10584-92
Miura K, Rus A, Sharkhuu A, Yokoi S, Karthikeyan AS, Raghothama KG, Baek D, Koo YD, Jin JB, Bressan RA, Yun DJ, Hasegawa PM. The Arabidopsis SUMO E3 ligase SIZ1 controls phosphate deficiency responses. Proc Natl Acad Sci U S A. 2005 May 24;102(21):7760-5
Sentis S, Le Romancer M, Bianchin C, Rostan MC, Corbo L. Sumoylation of the estrogen receptor alpha hinge region regulates its transcriptional activity. Mol Endocrinol. 2005 Nov;19(11):2671-84
Shuai K, Liu B. Regulation of gene-activation pathways by PIAS proteins in the immune system. Nat Rev Immunol. 2005 Aug;5(8):593-605
Levy C, Lee YN, Nechushtan H, Schueler-Furman O, Sonnenblick A, Hacohen S, Razin E. Identifying a common molecular mechanism for inhibition of MITF and STAT3 by PIAS3. Blood. 2006 Apr 1;107(7):2839-45
Ogata Y, Osaki T, Naka T, Iwahori K, Furukawa M, Nagatomo I, Kijima T, Kumagai T, Yoshida M, Tachibana I, Kawase I. Overexpression of PIAS3 suppresses cell growth and restores the drug sensitivity of human lung cancer cells in association with PI3-K/Akt inactivation. Neoplasia. 2006 Oct;8(10):817-25
Geiss-Friedlander R, Melchior F. Concepts in sumoylation: a decade on. Nat Rev Mol Cell Biol. 2007 Dec;8(12):947-56
Klopocki E, Schulze H, Strauss G, Ott CE, Hall J, Trotier F, Fleischhauer S, Greenhalgh L, Newbury-Ecob RA, Neumann LM, Habenicht R, König R, Seemanova E,
Megarbane A, Ropers HH, Ullmann R, Horn D, Mundlos S. Complex inheritance pattern resembling autosomal recessive inheritance involving a microdeletion in thrombocytopenia-absent radius syndrome. Am J Hum Genet. 2007 Feb;80(2):232-40
Martin S, Nishimune A, Mellor JR, Henley JM. SUMOylation regulates kainate-receptor-mediated synaptic transmission. Nature. 2007 May 17;447(7142):321-5
Palvimo JJ. PIAS proteins as regulators of small ubiquitin-related modifier (SUMO) modifications and transcription. Biochem Soc Trans. 2007 Dec;35(Pt 6):1405-8
Uzunova K, Göttsche K, Miteva M, Weisshaar SR, Glanemann C, Schnellhardt M, Niessen M, Scheel H, Hofmann K, Johnson ES, Praefcke GJ, Dohmen RJ. Ubiquitin-dependent proteolytic control of SUMO conjugates. J Biol Chem. 2007 Nov 23;282(47):34167-75
Brantley EC, Nabors LB, Gillespie GY, Choi YH, Palmer CA, Harrison K, Roarty K, Benveniste EN. Loss of protein inhibitors of activated STAT-3 expression in glioblastoma multiforme tumors: implications for STAT-3 activation and gene expression. Clin Cancer Res. 2008 Aug 1;14(15):4694-704
Kluge A, Dabir S, Kern J, Nethery D, Halmos B, Ma P, Dowlati A. Cooperative interaction between protein inhibitor of activated signal transducer and activator of transcription-3 with epidermal growth factor receptor blockade in lung cancer. Int J Cancer. 2009 Oct 1;125(7):1728-34
Onishi A, Peng GH, Hsu C, Alexis U, Chen S, Blackshaw S. Pias3-dependent SUMOylation directs rod photoreceptor development. Neuron. 2009 Jan 29;61(2):234-46
Rytinki MM, Kaikkonen S, Pehkonen P, Jääskeläinen T, Palvimo JJ. PIAS proteins: pleiotropic interactors associated with SUMO. Cell Mol Life Sci. 2009 Sep;66(18):3029-41
Spoden GA, Morandell D, Ehehalt D, Fiedler M, Jansen-Dürr P, Hermann M, Zwerschke W. The SUMO-E3 ligase PIAS3 targets pyruvate kinase M2. J Cell Biochem. 2009 May 15;107(2):293-302
Yagil Z, Kay G, Nechushtan H, Razin E. A specific epitope of protein inhibitor of activated STAT3 is responsible for the induction of apoptosis in rat transformed mast cells. J Immunol. 2009 Feb 15;182(4):2168-75
This article should be referenced as such:
Spoden G, Zwerschke W. PIAS3 (protein inhibitor of activated STAT, 3). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):38-41.
Gene Section Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 42
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
PSEN2 (presenilin 2 (Alzheimer disease 4)) Morgan Newman
School of Molecular and Biomedical Science, The University of Adelaide, Australia (MN)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/PSEN2ID41883ch1q42.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI PSEN2ID41883ch1q42.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity
Other names: AD3L, AD4, PS2, STM2
HGNC (Hugo): PSEN2
Location: 1q42.13
DNA/RNA
Description
Twelve exons, spans approximately 26.7 kb of
genomic DNA in the centromere to telomere
orientation, the translation initation codon is in
exon 4 and the stop codon in exon 12.
Transcription
mRNA of approximately 2.3 kb. Two alternatively
spliced transcript variants encoding different
isoforms of PSEN2 have been identified.
Pseudogene
Not known.
Protein
Description
The open reading frame encodes a 448 amino acid
protein, with an estimated molecular weight of 50
kDa.
It is a multi-spanning transmembrane protein with a
predicted 9 transmembrane domains.
Heterogeneous proteolytic processing generates N-
terminal and C-terminal fragments.
Expression
Neuronal (higher levels in hippocampus and
cerebellum). Isoform 1 is seen in the placenta,
skeletal muscle and heart while isoform 2 is seen in
the heart, brain, placenta, liver, skeletal muscle and
kidney. (In isoform 2 amino-acids 263-296 are
missing).
Localisation
Endoplasmic reticulum, plasma membrane, golgi
apparatus.
Function
Catalytic core of the gamma-secretase complex.
This complex catalyses the intramembrane cleavage
of single-pass membrane proteins such as Notch
and the Amyloid Precursor Protein (APP) to give
intracellular signaling. The released intracellular
domains of Notch or APP form complexes with
other proteins to regulate gene transcription.
Homology
The PSEN2 gene is conserved in chimpanzee, dog,
cow, mouse, rat, chicken, and zebrafish.
Presenilin 2 transcript, lines indicate introns and boxes exons. Untranslated regions are represented as yellow boxes and coding
regions as red boxes.
Presenilin 2 protein domains, bright blue boxes are transmembrane domains (TM).
PSEN2 (presenilin 2 (Alzheimer disease 4)) Newman M
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 43
Mutations
Somatic
23 mutations.
Nucleotide
change Disease Reference
Arg62His AD Cruts et al., 1998; Guerreiro et
al., 2008
Arg71Trp AD Guerreiro et al., 2008
Thr122Pro AD Finckh et al., 2000; 2005
Ser130Leu AD Sorbi et al., 2002; Tedde et al.,
2003; Tomaino et al., 2007
Val139Met AD Bernardi et al., 2008
Asn141Ile AD Levy-Lahad et al., 1995; Rogaev
et al., 1995
Met174Val AD Guerreiro et al., 2008
Ser175Cys AD Piscopo et al., 2008
Gln228Leu AD Zekanowski et al., 2003
Met239Ile AD Finckh et al., 2000
Met239Val AD Rogaev et al., 1995; Marcon et
al., 2004
Val393Met AD Lindquist et al., 2008; 2009
Thr430Met AD Lleo et al., 2002; Ezquerra et al.,
2003
Asp439Ala AD Lleo et al., 2001; 2002
Arg62His Breast
Cancer To et al., 2006
Arg71Trp Breast
Cancer To et al., 2006
Tyr231Cys FTD Marcon et al., 2008; 2009
Ala85Val LBD Piscopo et al., 2008
Thr122Arg Atypical
Dementia Binetti et al., 2003
Table. Mutations identified through genetic screening. AD: Alzheimer's Disease, FTD: Frontotemporal Dementia,
LBD: Lewy Body Dementia.
Implicated in
Breast cancer
Disease
Breast cancer is the most common form of cancer
for women. The cancer originates from the breast
tissue where it can be a ductal carcinoma or lobular
carcinoma. They can be further defined as in situ or
invasive cancers.
Oncogenesis
Mutations (see above).
Alzheimer's disease
Note
Mutations (see above) taken from the Alzheimer's
Disease and Frontotemporal Dementia Mutation
Database. Only pathogenic mutations are included.
Disease
Alzheimer's disease is the most prevalent form of
dementia. In affected individuals the disease causes
a progressive and permanent decline in memory and
cognitive abilities. Neuropathogenesis is proposed
to be a result of the accumulation of amyloid-beta
peptides in the brain together with increased
oxidative stress and neuroinflammation. The
presenilin proteins are central to the gamma-
secretase cleavage of the amyloid precursor protein
(APP), releasing the amyloid-beta peptide. Point
mutations in the presenilin genes lead to cases of
familial Alzheimer's disease (and some sporadic
cases) by altering APP cleavage resulting in excess
amyloid-beta formation.
Frontotemporal Dementia (FTD)
Note
Mutation (see above).
Disease
Frontotemporal dementia is a group of related
conditions resulting from the progressive
degeneration of the temporal and frontal lobes of
the brain (frontotemporal lobar degeneration,
FTLD), usually with the presence of abnormal
intracellular protein accumulations. These areas of
the brain play a significant role in decision-making,
behavioral control, emotion and language. The
disorder is often sporadic, familial FTD has been
linked to mutations in several genes, including
those encoding the microtubule-associated protein
tau (MAPT), progranulin (GRN), valosin-
containing protein (VCP) and charged
multivescicular body protein 2B (CHMP2B).
Lewy body Dementia (DLB)
Note
Mutation (see above).
Disease
Dementia with Lewy bodies is a neurodegenerative
disorder associated with abnormal structures (Lewy
bodies) which are clumps of alpha-synuclein and
ubiquitin protein in neurons found in certain areas
of the brain. In addition to dementia, patients with
dementia with Lewy bodies experience
hallucinations, motor impairment, and fluctuating
alertness.
PSEN2 (presenilin 2 (Alzheimer disease 4)) Newman M
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 44
Diagram taken from http://www.molgen.ua.ac.be/ADMutations. Coloured circles indicate mutation sites. Red: pathogenic,
orange: pathogenic nature unclear, green: not pathogenic.
To be noted
Note
Truncated variant PSEN2 protein (PS2V). Variant
transcript lacks exon 5 due to alternative splicing.
Encodes the first 119 codons of PSEN2 plus a
newly generated five amino acids SSMAG. PS2V is
detected in sporadic Alzheimer's disease, bi-polar
and schizophrenia cases (Sato et al., 1999; Smith et
al., 2004). Cell-culture experiments indicate that
this variant is upregulated under hypoxic conditions
(Sato et al., 1999).
References Levy-Lahad E, Wasco W, Poorkaj P, Romano DM, Oshima J, Pettingell WH, Yu CE, Jondro PD, Schmidt SD, Wang K. Candidate gene for the chromosome 1 familial Alzheimer's disease locus. Science. 1995 Aug 18;269(5226):973-7
Rogaev EI, Sherrington R, Rogaeva EA, Levesque G, Ikeda M, Liang Y, Chi H, Lin C, Holman K, Tsuda T. Familial Alzheimer's disease in kindreds with missense mutations in a gene on chromosome 1 related to the Alzheimer's disease type 3 gene. Nature. 1995 Aug 31;376(6543):775-8
Lee MK, Slunt HH, Martin LJ, Thinakaran G, Kim G, Gandy SE, Seeger M, Koo E, Price DL, Sisodia SS. Expression of presenilin 1 and 2 (PS1 and PS2) in human and murine tissues. J Neurosci. 1996 Dec 1;16(23):7513-25
Levy-Lahad E, Poorkaj P, Wang K, Fu YH, Oshima J, Mulligan J, Schellenberg GD. Genomic structure and expression of STM2, the chromosome 1 familial Alzheimer disease gene. Genomics. 1996 Jun 1;34(2):198-204
McMillan PJ, Leverenz JB, Poorkaj P, Schellenberg GD, Dorsa DM. Neuronal expression of STM2 mRNA in human brain is reduced in Alzheimer's disease. J Histochem Cytochem. 1996 Nov;44(11):1215-22
Prihar G, Fuldner RA, Perez-Tur J, Lincoln S, Duff K, Crook R, Hardy J, Philips CA, Venter C, Talbot C, Clark RF, Goate A, Li J, Potter H, Karran E, Roberts GW, Hutton M, Adams MD. Structure and alternative splicing of the presenilin-2 gene. Neuroreport. 1996 Jul 8;7(10):1680-4
Wolozin B, Iwasaki K, Vito P, Ganjei JK, Lacanà E, Sunderland T, Zhao B, Kusiak JW, Wasco W, D'Adamio L. Participation of presenilin 2 in apoptosis: enhanced basal activity conferred by an Alzheimer mutation. Science. 1996 Dec 6;274(5293):1710-3
Cruts M, Van Broeckhoven C. Presenilin mutations in Alzheimer's disease. Hum Mutat. 1998;11(3):183-90
Cruts M, Van Broeckhoven C. Molecular genetics of Alzheimer's disease. Ann Med. 1998 Dec;30(6):560-5
Cruts M, van Duijn CM, Backhovens H, Van den Broeck M, et al. Estimation of the genetic contribution of presenilin-1 and -2 mutations in a population-based study of presenile Alzheimer disease. Hum Mol Genet. 1998 Jan;7(1):43-51
Lao JI, Beyer K, Fernández-Novoa L, Cacabelos R. A novel mutation in the predicted TM2 domain of the presenilin 2 gene in a Spanish patient with late-onset Alzheimer's disease. Neurogenetics. 1998 Aug;1(4):293-6
PSEN2 (presenilin 2 (Alzheimer disease 4)) Newman M
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 45
Sato N, Hori O, Yamaguchi A, Lambert JC, Chartier-Harlin MC, Robinson PA, Delacourte A, Schmidt AM, Furuyama T, Imaizumi K, Tohyama M, Takagi T. A novel presenilin-2 splice variant in human Alzheimer's disease brain tissue. J Neurochem. 1999 Jun;72(6):2498-505
Finckh U, Alberici A, Antoniazzi M, Benussi L, Fedi V, Giannini C, Gal A, Nitsch RM, Binetti G. Variable expression of familial Alzheimer disease associated with presenilin 2 mutation M239I. Neurology. 2000 May 23;54(10):2006-8
Finckh U, Müller-Thomsen T, Mann U, Eggers C, Marksteiner J, Meins W, Binetti G, Alberici A, Hock C, Nitsch RM, Gal A. High prevalence of pathogenic mutations in patients with early-onset dementia detected by sequence analyses of four different genes. Am J Hum Genet. 2000 Jan;66(1):110-7
Lleó A, Blesa R, Gendre J, Castellví M, Pastor P, Queralt R, Oliva R. A novel presenilin 2 gene mutation (D439A) in a patient with early-onset Alzheimer's disease. Neurology. 2001 Nov 27;57(10):1926-8
Groth C, Nornes S, McCarty R, Tamme R, Lardelli M. Identification of a second presenilin gene in zebrafish with similarity to the human Alzheimer's disease gene presenilin2. Dev Genes Evol. 2002 Nov;212(10):486-90
Lleó A, Blesa R, Queralt R, Ezquerra M, Molinuevo JL, Peña-Casanova J, Rojo A, Oliva R. Frequency of mutations in the presenilin and amyloid precursor protein genes in early-onset Alzheimer disease in Spain. Arch Neurol. 2002 Nov;59(11):1759-63
Sorbi S, Tedde A, Nacmias B, Ciantelli M, Caffarra P, Ghidoni E, Bracco L, Piccini C.. Novel presenilin 1 and presenilin 2 mutations in early-onset Alzheimer's disease families. Neurobiology of Aging. 2002; 23(1S):S312.
Binetti G, Signorini S, Squitti R, Alberici A, Benussi L, Cassetta E, Frisoni GB, Barbiero L, Feudatari E, Nicosia F, Testa C, Zanetti O, Gennarelli M, Perani D, Anchisi D, Ghidoni R, Rossini PM. Atypical dementia associated with a novel presenilin-2 mutation. Ann Neurol. 2003 Dec;54(6):832-6
Ezquerra M, Lleó A, Castellví M, Queralt R, Santacruz P, Pastor P, Molinuevo JL, Blesa R, Oliva R. A novel mutation in the PSEN2 gene (T430M) associated with variable expression in a family with early-onset Alzheimer disease. Arch Neurol. 2003 Aug;60(8):1149-51
Nornes S, Groth C, Camp E, Ey P, Lardelli M. Developmental control of Presenilin1 expression, endoproteolysis, and interaction in zebrafish embryos. Exp Cell Res. 2003 Sep 10;289(1):124-32
Tedde A, Nacmias B, Ciantelli M, Forleo P, Cellini E, Bagnoli S, Piccini C, Caffarra P, Ghidoni E, Paganini M, Bracco L, Sorbi S. Identification of new presenilin gene mutations in early-onset familial Alzheimer disease. Arch Neurol. 2003 Nov;60(11):1541-4
Zekanowski C, Styczyńska M, Pepłońska B, Gabryelewicz T, Religa D, Ilkowski J, Kijanowska-Haładyna B, et al. Mutations in presenilin 1, presenilin 2 and amyloid precursor protein genes in patients with early-onset Alzheimer's disease in Poland. Exp Neurol. 2003 Dec;184(2):991-6
Marcon G, Giaccone G, Cupidi C, Balestrieri M, Beltrami CA, Finato N, Bergonzi P, Sorbi S, Bugiani O, Tagliavini F. Neuropathological and clinical phenotype of an Italian Alzheimer family with M239V mutation of presenilin 2 gene. J Neuropathol Exp Neurol. 2004 Mar;63(3):199-209
Smith MJ, Sharples RA, Evin G, McLean CA, Dean B, Pavey G, Fantino E, Cotton RG, Imaizumi K, Masters CL, Cappai R, Culvenor JG. Expression of truncated presenilin
2 splice variant in Alzheimer's disease, bipolar disorder, and schizophrenia brain cortex. Brain Res Mol Brain Res. 2004 Aug 23;127(1-2):128-35
Finckh U, Kuschel C, Anagnostouli M, Patsouris E, Pantes GV, Gatzonis S, Kapaki E, Davaki P, Lamszus K, Stavrou D, Gal A. Novel mutations and repeated findings of mutations in familial Alzheimer disease. Neurogenetics. 2005 May;6(2):85-9
To MD, Gokgoz N, Doyle TG, Donoviel DB, Knight JA, Hyslop PS, Bernstein A, Andrulis IL. Functional characterization of novel presenilin-2 variants identified in human breast cancers. Oncogene. 2006 Jun 15;25(25):3557-64
Tomaino C, Bernardi L, Anfossi M, Costanzo A, Ferrise F, Gallo M, Geracitano S, Maletta R, Curcio SA, Mirabelli M, Colao R, Frangipane F, Puccio G, Calignano C, Muraca MG, Paonessa A, Smirne N, Leotta A, Bruni AC. Presenilin 2 Ser130Leu mutation in a case of late-onset "sporadic" Alzheimer's disease. J Neurol. 2007 Mar;254(3):391-3
Bernardi L, Tomaino C, Anfossi M, Gallo M, Geracitano S, Puccio G, Colao R, Frangipane F, Mirabelli M, Smirne N, Giovanni Maletta R, Bruni AC. Late onset familial Alzheimer's disease: novel presenilin 2 mutation and PS1 E318G polymorphism. J Neurol. 2008 Apr;255(4):604-6
Brouwers N, Sleegers K, Van Broeckhoven C. Molecular genetics of Alzheimer's disease: an update. Ann Med. 2008;40(8):562-83
Lindquist SG, Hasholt L, Bahl JM, Heegaard NH, Andersen BB, Nørremølle A, Stokholm J, Schwartz M, Batbayli M, Laursen H, Pardossi-Piquard R, Chen F, St George-Hyslop P, Waldemar G, Nielsen JE. A novel presenilin 2 mutation (V393M) in early-onset dementia with profound language impairment. Eur J Neurol. 2008 Oct;15(10):1135-9
Nornes S, Newman M, Verdile G, Wells S, Stoick-Cooper CL, Tucker B, Frederich-Sleptsova I, Martins R, Lardelli M. Interference with splicing of Presenilin transcripts has potent dominant negative effects on Presenilin activity. Hum Mol Genet. 2008 Feb 1;17(3):402-12
Piscopo P, Marcon G, Piras MR, Crestini A, Campeggi LM, Deiana E, Cherchi R, Tanda F, Deplano A, Vanacore N, Tagliavini F, Pocchiari M, Giaccone G, Confaloni A. A novel PSEN2 mutation associated with a peculiar phenotype. Neurology. 2008 Apr 22;70(17):1549-54
Lindquist SG, Schwartz M, Batbayli M, Waldemar G, Nielsen JE. Genetic testing in familial AD and FTD: mutation and phenotype spectrum in a Danish cohort. Clin Genet. 2009 Aug;76(2):205-9
Marcon G, Di Fede G, Giaccone G, Rossi G, Giovagnoli AR, Maccagnano E, Tagliavini F. A novel Italian presenilin 2 gene mutation with prevalent behavioral phenotype. J Alzheimers Dis. 2009;16(3):509-11
Nornes S, Newman M, Wells S, Verdile G, Martins RN, Lardelli M. Independent and cooperative action of Psen2 with Psen1 in zebrafish embryos. Exp Cell Res. 2009 Oct 1;315(16):2791-801
Guerreiro RJ, Baquero M, Blesa R, Boada M, Brás JM, et al. Genetic screening of Alzheimer's disease genes in Iberian and African samples yields novel mutations in presenilins and APP. Neurobiol Aging. 2010 May;31(5):725-31
This article should be referenced as such:
Newman M. PSEN2 (presenilin 2 (Alzheimer disease 4)). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):42-45.
Gene Section Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 46
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
RASSF6 (Ras association (RalGDS/AF-6) domain family member 6) Luke B Hesson, Farida Latif
Lowy Cancer Centre and Prince of Wales Clinical School, Faculty of Medicine, University of New
South Wales, NSW2052, Australia (LBH), School of Clinical and Experimental Medicine, College of
Medical and Dental Sciences, Department of Medical and Molecular Genetics, University of
Birmingham, Birmingham B15 2TT, UK (FL)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/RASSF6ID43462ch4q13.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI RASSF6ID43462ch4q13.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity
Other names: DKFZp686K23225
HGNC (Hugo): RASSF6
Location: 4q13.3
Local order: Centromere-AFP-AFM-RASSF6-
IL8-Telomere.
Note
Brief overview : The RASSF family of tumour
suppressor genes (TSG) encode Ras superfamily
effector proteins that, amongst other functions,
mediate some of the growth inhibitory functions of
Ras proteins. Several members of this family are
inactivated by promoter DNA hypermethylation in
a broad range of cancers, and the RASSF6 gene
may be a frequent target of epigenetic inactivation
in leukaemias. The RASSF6 protein is involved in
the regulation of apoptosis partly by controlling the
function of the proapoptotic mammalian
serine/threonine kinases 1 and 2 (MST1 and MST2)
and modulator of apoptosis 1 (MOAP-1).
Figure 1: RASSF6 gene structure. The RASSF6 gene is composed of at least two isoforms that are transcribed from
immediately upstream (RASSF6A) or from within (RASSF6B) a small CpG island.
RASSF6 (Ras association (RalGDS/AF-6) domain family member 6)
Hesson LB, Latif F
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 47
DNA/RNA
Description
The RASSF6 gene occupies 47478 bp of genomic
DNA. RASSF6A [GenBank:NM_201431] contains
11 exons and is transcribed 82 bp upstream from a
small (214 bp) 5' CpG island (at chr4:74,486,045-
74,486,258). RASSF6B [GenBank:NM_177532]
contains 11 exons and is transcribed from within
the same CpG island. There is evidence of
additional splice variants and transcription initiation
sites for the RASSF6 gene, however these have not
been validated.
Protein
Description
RASSF6A [GenBank:NP_958834] is a 369 amino
acid protein whereas RASSF6B
[GenBank:NP_803876] is a 337 amino acid protein.
The RASSF6A protein (figure 2) contains C-
terminal Ras-association (RA) and
Sav/RASSF/Hpo (SARAH) domains that define the
'classical' RASSF family (RASSF1, RASSF2,
RASSF3, RASSF4, RASSF5, RASSF6). The
RASSF6B protein lacks the N-terminal 32 amino
acids present in RASSF6A but is otherwise
identical. There are some inconsistencies in the
literature regarding which may be the major
isoform, however all functional work to date has
studied the larger 369 amino acid protein, which is
hereafter referred to as RASSF6. Using an in-house
antibody Ikeda et al., (2007) were unable to detect
endogenous RASSF6 protein. All functional work
has been performed using overexpressed RASSF6
protein. A commercially available antibody towards
RASSF6 (ProteinTech Group) has been used to
demonstrate re-expression following treatment with
the DNA demethylating agent 5-aza-
2'deoxycytidine (5azaDC) and Trichostatin A
(TSA) in leukaemia cell lines in which RASSF6 is
epigenetically inactivated (Hesson et al., 2009).
Expression
Northern blotting of a normal tissue RNA panel
shows RASSF6 mRNA is highly expressed in
thymus, kidney and placenta, with lower levels of
expression in colon, small intestine and lung (Allen
et al., 2007). In the same study the matched normal
tissue from a primary tumour cDNA panel showed
readily detectable levels of RASSF6 expression in
rectal, pancreatic, liver, breast and stomach tissues.
There has been no systematic analysis of the
expression patterns of the different RASSF6
isoforms. RT-PCR analysis of a range of tumour
cell lines showed that RASSF6 is highly expressed
in HeLa and A549 cells with lower levels observed
in MCF-7, U373, H1299 and HepG2 (Ikeda et al.,
2007). It appears from these studies that the tissue
distribution of RASSF6 expression is much more
restricted than that of the RASSF members
RASSF1, RASSF2 and RASSF5.
Expression of the RASSF6 gene is lost or
downregulated in a variety of solid tumours by
unknown mechanisms, whereas in childhood
leukaemias RASSF6 is inactivated by CpG island
promoter region DNA methylation (see below).
Function
dRASSF antagonises the Hippo pathway
In Drosophila, dRASSF represents the orthologue
of mammalian RASSF1-6. dRASSF protein
competes with Salvador (the Drosophila orthologue
of the mammalian WW45 protein) for binding to
Hippo (the Drosophila orthologue of the
mammalian MST kinases). The dRASSF-Hippo
and Salvador-Hippo interactions appear mutually
exclusive and control Hippo function in very
different ways.
Figure 2: RASSF6 transcript and protein structure. The RASSF6A mRNA (red bar) encodes a 369 amino acid protein
[GenBank:NP_958834] containing a C-terminal Ras-association (RA) domain of the RalGDS/AF-6 variety and acidic coiled-coil Sav/RASSF/Hpo (SARAH) domain. RASSF6B [GenBank:NP_803876] is a predicted 337 amino acid protein lacking the N-
terminal 32 amino acids present in RASSF6A.
RASSF6 (Ras association (RalGDS/AF-6) domain family member 6)
Hesson LB, Latif F
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 48
Both Salvador or dRASSF are stabilised following
binding to Hippo, however, probing of
immunoprecipitates of Salvador or dRASSF with a
phospho-specific antibody that recognises active
Hippo demonstrates that Hippo is present in
Drosophila cells in two pools; an active form
associated with Salvador and an inactive form
associated with dRASSF (Polesello et al., 2006).
Furthermore, RNAi-mediated dRASSF depletion
led to a marked increase in Hippo activation
following Staurosporine (STS) treatment, a potent
activator of Hippo in Drosophila cells and MSTs in
mammalian cells. Thus, in Drosophila dRASSF
restricts the activation of Hippo, which may
account for the smaller size of dRASSF mutant flies
(Polesello et al., 2006).
Regulation of apoptosis by mammalian RASSF6
Several studies have demonstrated that
overexpression of RASSF6 induces apoptosis and
inhibits the growth of a variety of tumour cell lines
(Ikeda et al., 2007; Allen et al., 2007; Ikeda et al.,
2009). In HeLa cells RASSF6-induced apoptosis
occurred through both caspase-dependent and
caspase-independent pathways, since
overexpression of RASSF6 results in cleavage and
activation of caspase-3 but apoptosis was not
abrogated by z-VAD-FMK, an inhibitor of caspase-
1, caspase-3, caspase-4 and caspase-7 activation
(Ikeda et al., 2007). RASSF6 also induced Bax
activation and cytochrome C release as well as the
release of apoptosis-inducing factor (AIF) and
endonuclease G (endoG) from the mitochondria.
Following release from the mitochondria AIF and
endoG may result in DNA fragmentation even in
the absence of caspase activation. Early evidence
has suggested that the molecular mechanisms of
RASSF6-induced apoptosis are likely to be
complex and multi-layered. Currently, three
signalling routes are implicated in RASSF6-
induced apoptosis (see below), though at present it
is unclear whether these routes act autonomously or
as part of an extensive apoptotic network.
RASSF6 is an effector molecule of K-Ras-
mediated apoptosis
RASSF6 interacts with K-Ras in a GTP-dependent
manner via its Ras-association domain and the
effector domain of K-Ras. Therefore, RASSF6
exhibits the basic properties of a Ras effector
protein. These data are in contradiction to another
report in which RASSF6 did not interact with K-
Ras, H-Ras, N-Ras, M-Ras or TC21 under the same
conditions that RASSF5 binds to these Ras proteins
(Ikeda et al., 2007). Apart from a strict context-
dependency of these interactions The reasons for
this discrepancy is unclear but may be related to a
strict context-dependency of the interaction or to
the requirement of Ras farnesylation as shown by
Allen et al., (2007). Of particular note however, is
the observation that RASSF6 acts synergistically
with activated K-Ras to induce cell death in 293-T
cells (Allen et al., 2007). Taken together these data
suggests RASSF6 does indeed function within a K-
Ras-regulated pathway, most likely through direct
interaction, to determine cell fate.
RASSF6 negatively regulates the proapoptotic
protein MST2
RASSF6 interacts with MST2 via the
Sav/RASSF/Hpo (SARAH) domains within both
proteins (Ikeda et al., 2009). It seems clear from
many studies that several (if not all) classical
RASSF proteins interact with the MST1 and MST2
kinases and that this interaction is at least partly
involved in RASSF-induced apoptosis (van der
Weyden and Adams, 2007). Ikeda et al., (2009)
showed that the SARAH domain of MST2 can bind
both WW45 (the mammalian orthologue of the
Drosophila protein Salvador) and RASSF6 to form
a trimeric complex. However, RASSF6 and WW45
do not interact. This differs somewhat with the
regulation of the Hippo pathway in Drosophila, in
which the Salvador/Hippo and dRASSF/Hippo
complexes are mutually exclusive (see above).
RASSF6 inhibits MST2 kinase activity, however
activation of MST2 releases RASSF6 in a manner
dependent on WW45. Previous studies have
demonstrated that MST2 can extensively
phosphorylate WW45 (Callus et al., 2006). Ikeda et
al., (2009) demonstrated that activation of MST2 by
the phosphatase inhibitor okadaic acid (OA)
reduced the co-immunoprecipitation of RASSF6
with MST2, whilst the association of MST2 and
WW45 remained unchanged. Taken together this
suggests that the activation of MST2 and
subsequent phosphorylation of WW45 results in the
release of RASSF6 from the
WW45/MST2/RASSF6 complex. The release of
RASSF6 was shown to be WW45-dependent since
RASSF6 remained in complex with MST2
following OA treatment of cells in which WW45
had been depleted by RNAi. Furthermore, the
release of RASSF6 appears to be necessary for
RASSF6-mediated apoptosis. However, apoptosis
was vastly reduced in cells transfected with both
MST2 and RASSF6. This block of RASSF6-
induced apoptosis was dependent on the SARAH
domain of MST2 but not the kinase activity of
MST2 suggesting that MST2 blocks RASSF6-
mediated apoptosis by physical interaction.
Interestingly co-expression of RASSF6, MST2 and
WW45 resulted in potent cell death. This effect was
dependent on the SARAH domain of WW45
suggesting that physical interaction of WW45 with
MST2 was necessary to reinstate RASSF6-induced
apoptosis by allowing the release of RASSF6.
Inhibition of MST2 kinase activity by RASSF6 also
results in the inhibition of NDR1 and LATS2
phosphorylation (Ikeda et al., 2009). NDR1 and
LATS2 are known MST2 substrates that form part
of the MST/Hippo tumour suppressor pathway (see
Hergovich and Hemmings, 2009 and Harvey and
RASSF6 (Ras association (RalGDS/AF-6) domain family member 6)
Hesson LB, Latif F
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 49
Tapon, 2007 for a more thorough review of the
regulation of apoptosis through the MST/Hippo
pathway). Thus release of RASSF6 from the
MST2-WW45 complex allows RASSF6-mediated
and MST2-mediated apoptosis (figure 3). The
induction of apoptosis through these separate
pathways may explain the caspase-dependent and
caspase-independent nature of RASSF6-mediated
apoptosis, since full activation of MSTs is thought
to require caspase cleavage.
MOAP-1 is involved in RASSF6-mediated
apoptosis
The MOAP-1 protein may also be a RASSF6
effector molecule. To date, two independent studies
have demonstrated that RASSF6 also interacts with
MOAP-1 (Allen et al., 2007; Ikeda et al., 2009).
Though this interaction has not been demonstrated
formally at the endogenous level, co-expression of
the two proteins clearly demonstrates that MOAP-1
is a likely mediator of RASSF6-induced apoptosis
in a manner independent of the MST/Hippo tumour
suppressor pathway. MOAP-1 regulates the
'extrinsic' pathway of apoptosis by acting
downstream of death receptors such as the tumour
necrosis factor a receptor 1 (TNF-R1) and TNFa
apoptosis-inducing related ligand receptor 1
(TRAIL-R1). A role for MOAP-1 in regulating
RASSF1A-induced apoptosis has previously been
demonstrated. Following ligand binding the C-
terminal region of MOAP-1 associates with the
death domain of TNF-R1. Subsequently, the TNF-
R1/MOAP-1 receptor complex is internalised and
recruits RASSF1A through the N-terminal cysteine-
rich (C1) domain within RASSF1A (Baksh et al.,
2005; Vos et al., 2006; Foley et al., 2008). Under
'static' conditions MOAP-1 is held in an inactive
conformation, however binding to RASSF1A
results in a conformational change that allows
MOAP-1 to interact with Bax. This in turn induces
a conformational change within Bax that is required
for its insertion into the mitochondrial membrane
and for the release of inner mitochondrial
membrane proteins that can induce apoptosis.
Interestingly, activated K-Ras, RASSF1A and
MOAP-1 synergise to induce Bax activation and
apoptosis (Vos et al., 2006) indicating that cell
death induced by the RASSF1A/MOAP-1
interaction may be regulated by both K-Ras and
death receptors.
The acidic sequence Glu-Glu-Glu-Glu [312
EEEE] in
the SARAH domain of RASSF1A that binds to
MOAP-1 (Baksh et al., 2005) is also partially
conserved in the RASSF6 SARAH domain
[EEEK]. However, this is unlikely to be the sole
site of interaction since RASSF6 lacking the N-
terminal region, the Ras-association domain, or the
SARAH domain also interacted with MOAP-1
(Ikeda et al., 2009). Depletion of MOAP-1 partially
suppresses RASSF6-mediated apoptosis and co-
expression of MST2 vastly reduces the co-
immunoprecipitation of RASSF6 and MOAP-1; an
effect that is abrogated by the expression of WW45.
Taken together these data suggest that the
MST2/WW45 complex binds RASSF6 and
prevents its interaction with MOAP-1. The MST-
Hippo pathway and the MOAP-1 pathway represent
distinct RASSF6-regulated apoptotic pathways that
are triggered by the activation of MST2 (figure 3).
Homology
RASSF6 is one of 10 members of the Ras-
association domain family (RASSF) comprising
RASSF1-10 (please refer to figure 3 of the RASSF2
gene card. RASSF1-6 are termed the 'classical'
RASSF family and contain C-terminal RA and
SARAH domains. Consequently, RASSF1-6 are
most similar in sequence within their C-termini.
RASSF7, RASSF8, RASSF9 and RASSF10
represent evolutionarily conserved but structurally
distinct RASSF members that lack the SARAH
domains and contain N-terminal RA domains.
RASSF7-10 are termed the 'N-terminal' RASSF
family. Many of these RASSF members are
involved in tumourigenesis and several (RASSF1A,
RASSF2, RASSF4, RASSF5A, RASSF6 and
RASSF10) are inactivated in a variety of human
cancers (Hesson et al., 2007; van der Weyden and
Adams, 2007; Hesson et al., 2009). RASSF6 also
has orthologues in several species (table 1)
including Drosophila melanogaster, which is the
orthologue of the human RASSF1-6 genes.
RASSF6 (Ras association (RalGDS/AF-6) domain family member 6)
Hesson LB, Latif F
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 50
Figure 3: The molecular basis of RASSF6-induced apoptosis. Multiple lines of evidence suggest that RASSF6-induced
apoptosis involves multiple pathways. K-Ras, MST2 and MOAP-1 have all been implicated in RASSF6-induced apoptosis. To date RASSF1, RASSF2, RASSF4, RASSF5 and RASSF6 have been shown to bind to the MST1 and MST2 kinases. Thus, the
RASSF proteins play a major role in regulating apoptosis through the MST/Hippo tumour suppressor pathway leading to apoptosis. The regulation of apoptosis through activation of the TNF-R1 and TRAIL-R1 death receptor pathways has been
shown to involve the interaction of RASSF1A with MOAP-1. This interaction leads to the release of inner mitochondrial membrane proteins that result in apoptosis. RASSF6 also interacts with MOAP-1, though the events downstream have not been fully elucidated. The release of inner mitochondrial membrane proteins is a major facet of RASSF6-induced apoptosis therefore it
seems likely that RASSF6 regulates MOAP-1 function in a similar way to RASSF1A and may feed into the same pathway. The synergistic effects of activated K-Ras, RASSF6 and MOAP-1 overexpression suggests that apoptosis through the
RASSF6/MOAP-1 complex may be at least partially K-Ras-regulated.
RASSF6 (Ras association (RalGDS/AF-6) domain family member 6)
Hesson LB, Latif F
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 51
Table 1: RASSF6 orthologues in model species.
Mutations
Note
To date no inactivating mutations to RASSF6 have
been described.
Implicated in
Various cancers
Note
Loss of RASSF6 expression in cancer.
Disease
Northern blotting analysis showed RASSF6
expression is lost or downregulated in 30-60% of
primary tumour tissues of the breast, colon, kidney,
liver, pancreas, stomach and thyroid (Allen et al.,
2007). The mechanisms underlying this reduced
expression are not clear and, at least in solid
tumours, loss of RASSF6 expression is not
associated with promoter DNA methylation (Allen
et al., 2007). Other mechanisms of gene silencing
such as histone modifications or long-range
epigenetic silencing may account for RASSF6
downregulation, but this has not been investigated.
In childhood leukaemias silencing of RASSF6
expression by promoter hypermethylation appears
to be an extremely frequent event and was
identified in 94% (48/51) B-ALL and 41% (12/29)
T-ALL (Hesson et al., 2009). To date this remains
the first and only description of a mechanism
accounting for the loss of RASSF6 expression in
cancer but further suggests that epigenetic
inactivation of RASSF6 may be specific to
leukaemias. Further evidence that the loss of
RASSF6 expression is important in cancer is
provided in a study by Finn et al., (2007), which
demonstrated downregulation of RASSF6
expression in malignant versus benign thyroid
tissue.
Pancreatic endocrine tumour
Note
RASSF6 is downregulated in the pancreatic
endocrine tumour cell line BON1 following siRNAi
of Achaete-scute complex-like 1 (ASCL1), a basic
helix loop helix (bHLH) transcription factor
regulated by the NOTCH signalling pathway
RASSF6 (Ras association (RalGDS/AF-6) domain family member 6)
Hesson LB, Latif F
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 52
(Johansson et al., 2009). ASCL1 also negatively
regulates expression of Dickkopf homologue 1
(DKK1), an antagonist of the Wnt/beta-catenin-
dependent signalling pathway.
Breast cancer
Note
Using an in vitro model of breast carcinogenesis,
human non-tumourigenic immortalised breast
epithelial cell line MCF-10A cells were selected
following exposure to the mutagen ICR191 to
create transformed cells (Zientek-Targosz et al.,
2008). One of the genes found to contain a
frameshift mutation following this selection was
RASSF6. The authors postulate that this indicates
that inactivation of RASSF6 may be involved in the
transformation of breast epithelial cells thus
indicating the potential importance of RASSF6 in
tumour development.
Childhood B-ALL
Note
RASSF6 is downregulated in mature B-cells in
response to B-cell activating factor (BAFF)
treatment (Saito et al., 2008). BAFF is a tumour
necrosis factor (TNF) superfamily member, which
is thought to be involved in the survival and
maturation of B-cells partly by inhibiting apoptosis.
This finding reiterates the potential importance of
the frequent epigenetic inactivation of RASSF6 in
childhood B-ALL (Hesson et al., 2009), which
would nullify RASSF6-mediated apoptosis.
Viral-induced bronchiolitis
Disease
Genetic mapping studies have linked the RASSF6
locus to viral-induced bronchiolitis, the most
common cause of infant hospital admissions in the
industrialised world (Hull et al., 2004; Smyth and
Openshaw, 2006). The 250 kb interval at 4q13.3
implicated by Hull et al., (2004) includes three
genes (AFM, RASSF6 and IL8), however it is
unclear which gene is the important player. The
predominant cause of acute viral bronchiolitis is
infection with the respiratory syncytial virus (RSV).
A major consequence of RSV infection is
stimulation of the NF-kappaB pathway (Hull et al.,
2004), which is thought to modulate the degree of
inflammation and support viral replication, perhaps
by suppressing apoptosis (Bitko et al., 2004). Using
an NF-kappaB luciferase reporter system in A549
lung tumour cells it has been found that RASSF6
can suppress the serum-induced basal levels of NF-
kappaB reporter expression by approximately
fivefold (Allen et al., 2007). Therefore, RASSF6
remains a promising susceptibility gene candidate
for viral-induced bronchiolitis.
Heavy metal detoxification
Note
Expression of murine Rassf6 was upregulated
following subcutaneous injection of Cadmium,
possibly implicating Rassf6 in the cellular
mechanisms of heavy metal detoxification
(Wimmer et al., 2005). However, it seems equally
likely that the observed upregulation of Rassf6
could be due to increased apoptosis caused by the
toxic effects of heavy metal exposure.
References Bitko V, Garmon NE, Cao T, Estrada B, Oakes JE, Lausch RN, Barik S. Activation of cytokines and NF-kappa B in corneal epithelial cells infected by respiratory syncytial virus: potential relevance in ocular inflammation and respiratory infection. BMC Microbiol. 2004 Jul 15;4:28
Hull J, Rowlands K, Lockhart E, Sharland M, Moore C, Hanchard N, Kwiatkowski DP. Haplotype mapping of the bronchiolitis susceptibility locus near IL8. Hum Genet. 2004 Feb;114(3):272-9
Baksh S, Tommasi S, Fenton S, Yu VC, Martins LM, Pfeifer GP, Latif F, Downward J, Neel BG. The tumor suppressor RASSF1A and MAP-1 link death receptor signaling to Bax conformational change and cell death. Mol Cell. 2005 Jun 10;18(6):637-50
Wimmer U, Wang Y, Georgiev O, Schaffner W. Two major branches of anti-cadmium defense in the mouse: MTF-1/metallothioneins and glutathione. Nucleic Acids Res. 2005;33(18):5715-27
Callus BA, Verhagen AM, Vaux DL. Association of mammalian sterile twenty kinases, Mst1 and Mst2, with hSalvador via C-terminal coiled-coil domains, leads to its stabilization and phosphorylation. FEBS J. 2006 Sep;273(18):4264-76
Polesello C, Huelsmann S, Brown NH, Tapon N. The Drosophila RASSF homolog antagonizes the hippo pathway. Curr Biol. 2006 Dec 19;16(24):2459-65
Smyth RL, Openshaw PJ. Bronchiolitis. Lancet. 2006 Jul 22;368(9532):312-22
Vos MD, Dallol A, Eckfeld K, Allen NP, Donninger H, Hesson LB, Calvisi D, Latif F, Clark GJ. The RASSF1A tumor suppressor activates Bax via MOAP-1. J Biol Chem. 2006 Feb 24;281(8):4557-63
Allen NP, Donninger H, Vos MD, Eckfeld K, Hesson L, Gordon L, Birrer MJ, Latif F, Clark GJ. RASSF6 is a novel member of the RASSF family of tumor suppressors. Oncogene. 2007 Sep 13;26(42):6203-11
Finn SP, Smyth P, Cahill S, Streck C, O'Regan EM, Flavin R, Sherlock J, Howells D, Henfrey R, Cullen M, Toner M, Timon C, O'Leary JJ, Sheils OM. Expression microarray analysis of papillary thyroid carcinoma and benign thyroid tissue: emphasis on the follicular variant and potential markers of malignancy. Virchows Arch. 2007 Mar;450(3):249-60
Harvey K, Tapon N. The Salvador-Warts-Hippo pathway - an emerging tumour-suppressor network. Nat Rev Cancer. 2007 Mar;7(3):182-91
Hesson LB, Cooper WN, Latif F. The role of RASSF1A methylation in cancer. Dis Markers. 2007;23(1-2):73-87
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Ikeda M, Hirabayashi S, Fujiwara N, Mori H, Kawata A, Iida J, Bao Y, Sato Y, Iida T, Sugimura H, Hata Y. Ras-association domain family protein 6 induces apoptosis via both caspase-dependent and caspase-independent pathways. Exp Cell Res. 2007 Apr 15;313(7):1484-95
van der Weyden L, Adams DJ. The Ras-association domain family (RASSF) members and their role in human tumourigenesis. Biochim Biophys Acta. 2007 Sep;1776(1):58-85
Foley CJ, Freedman H, Choo SL, Onyskiw C, Fu NY, Yu VC, Tuszynski J, Pratt JC, Baksh S. Dynamics of RASSF1A/MOAP-1 association with death receptors. Mol Cell Biol. 2008 Jul;28(14):4520-35
Saito Y, Miyagawa Y, Onda K, Nakajima H, Sato B, Horiuchi Y, Okita H, Katagiri YU, Saito M, Shimizu T, Fujimoto J, Kiyokawa N. B-cell-activating factor inhibits CD20-mediated and B-cell receptor-mediated apoptosis in human B cells. Immunology. 2008 Dec;125(4):570-90
Zientek-Targosz H, Kunnev D, Hawthorn L, Venkov M, Matsui S, Cheney RT, Ionov Y. Transformation of MCF-10A cells by random mutagenesis with frameshift mutagen ICR191: a model for identifying candidate breast-tumor suppressors. Mol Cancer. 2008 Jun 5;7:51
Hergovich A, Hemmings BA. Mammalian NDR/LATS protein kinases in hippo tumor suppressor signaling. Biofactors. 2009 Jul-Aug;35(4):338-45
Hesson LB, Dunwell TL, Cooper WN, Catchpoole D, Brini AT, Chiaramonte R, Griffiths M, Chalmers AD, Maher ER, Latif F. The novel RASSF6 and RASSF10 candidate tumour suppressor genes are frequently epigenetically inactivated in childhood leukaemias. Mol Cancer. 2009 Jul 1;8:42
Ikeda M, Kawata A, Nishikawa M, Tateishi Y, Yamaguchi M, Nakagawa K, Hirabayashi S, Bao Y, Hidaka S, Hirata Y, Hata Y. Hippo pathway-dependent and -independent roles of RASSF6. Sci Signal. 2009 Sep 29;2(90):ra59
Johansson TA, Westin G, Skogseid B. Identification of Achaete-scute complex-like 1 (ASCL1) target genes and evaluation of DKK1 and TPH1 expression in pancreatic endocrine tumours. BMC Cancer. 2009 Sep 10;9:321
This article should be referenced as such:
Hesson LB, Latif F. RASSF6 (Ras association (RalGDS/AF-6) domain family member 6). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):46-53.
Gene Section Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 54
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
RPA2 (replication protein A2, 32kDa) Anar KZ Murphy, James A Borowiec
Dept of Biochemistry and New York University Cancer Institute, New York University School of
Medicine, New York, New York 10016, USA (AKZM, JAB)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/RPA2ID42146ch1p35.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI RPA2ID42146ch1p35.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity
Other names: REPA2, RPA32
HGNC (Hugo): RPA2
Location: 1p35.3
Local order: The human RPA2 gene maps on
1p35.3 between the SMPDL3B (sphingomyelin
phosphodiesterase, acid-like 3B) and C1orf38
(interferon-gamma inducible gene ICB-1 (induced
by contact to basement membrane)).
DNA/RNA
Description
The RPA2 gene is contained within 24.5 kb of
chromosome 1.
The coding sequence is contained within nine
exons. There is no confirmed alternative splicing of
the RPA2 gene, or differential promoter usage.
Transcription
The RPA2 mRNA transcript is 1.5 kb. The RPA2
promoter contains four E2F consensus sequences
within the region about 400 bp upstream of the
mRNA start site, and putative binding sites for
ATF-1 and SP-1 transcription factors. Expression is
upregulated 2 to 3-fold by E2F, with mutation of
the three start site-proximal E2F sites causing a loss
of E2F responsiveness (Kalma et al., 2001).
Pseudogene
RPA2 does not have known pseudogenes.
The sequence numbering corresponds to EMBL locus DQ001128 (26.6 kb). Exon are indicated as boxes (yellow = 5' UTR, blue = CDS, red = 3' UTR), and introns with orange lines. Two lengthy introns have been truncated (indicated with parallel diagonal
lines) to improve viewability.
RPA2 (replication protein A2, 32kDa) Murphy AKZ, Borowiec JA
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 55
Protein
Upper panel: Schematic showing the key domains of RPA2.
Lower panel: RPA2 phosphorylation sites are shown in bold, with the primary responsible kinases indicated above each site. Some sites can be phosphorylated by more than
one kinase (e.g., T21 by ATM and DNA-PK).
Description
RPA2 is the middle subunit of the heterotrimeric
Replication Protein A (RPA; (reviewed in Binz et
al., 2004)). The subunit is composed of 270
residues, and has a nominal molecular weight of
29.2 kDa. RPA2 contains an N-terminal
phosphorylation region with 7 phosphorylation
sites, a central DNA-binding domain (termed DBD-
D), and a C-terminal region that can form a three-
helix bundle. One helix of the three helix bundle is
contributed by each RPA subunit, with this
structure responsible for supporting
heterotrimerization of the RPA complex
(Bochkareva et al., 2002). At least in the non-
phosphorylated state, the N-terminal region is
unstructured. DBD-D is constructed from an
oligonucleotide/oligosaccharide binding (OB) fold
(Bochkarev et al., 1999), one of six OB folds found
with the RPA heterotrimer (four OB folds are
located in RPA1, and one within RPA3).
Expression
RPA is an essential factor for DNA replication and
repair, and hence is expressed in all tissues.
Localisation
Nuclear.
Function
General function: RPA is a heterotrimeric single-
stranded DNA (ssDNA) binding protein that is
essential for chromosomal DNA replication,
homologous recombination, and particular DNA
repair reactions (nucleotide excision repair). The
apparent association constant of the RPA: ssDNA
complex is 109 - 10
11 M
-1 (Kim et al., 1992). While
RPA2 contains a central DBD (Philipova et al.,
1996), the major effect of mutating DBD-D is to
decrease the size of the ssDNA occluded by RPA
binding, with only minor effects on RPA: ssDNA
affinity (Bastin-Shanower and Brill, 2001). A key
function of the RPA2 subunit is to regulate RPA
activity in DNA replication and repair reactions,
through the RPA2 phosphorylation state (see
below).
1) RPA2 phosphorylation. The N-terminal 33
residues of RPA2 contain seven phosphorylation
sites. In interphase cells, genotoxic stress (e.g.,
caused by chromosomal double-strand DNA breaks
or DNA replication stress) induces RPA2
phosphorylation by members of the
phosphatidylinositol 3-kinase-like kinase (PIKK;
ATM, ATR, and DNA-PK) and cyclin-dependent
kinases (CDK) families (reviewed in Binz et al.,
2004). Mutation of particular RPA2
phosphorylation sites causes defects in homologous
recombination (Lee et al., 2010), and Rad51
recruitment to nuclear repair foci (Anantha et al.,
2008; Lee et al., 2010). Mutation of these sites also
causes genomic instability in response to DNA
replication stress induced by cellular treatment with
hydroxyurea (Vassin et al., 2009). RPA
phosphorylation also increases cell viability in
response to DNA damage arising during mitosis
(Anantha et al., 2008). Modification of sites in the
phosphorylation region of RPA2 proceeds in a
favored order in response to genotoxic stress
(Anantha et al., 2007). The phosphorylation of
individual RPA2 residues is dependent on the type
of DNA damage or replication stress encountered
(Anantha et al., 2007; Vassin et al., 2009). RPA2 is
a substrate both for PP2A and PP4 phosphatases
(Feng et al., 2009; Lee et al., 2010).
2) Involvement of RPA2 in protein-protein
interactions. RPA2 interacts with the nucleotide
excision repair factor XPA (He et al., 1995), base
excision repair enzyme UNG2 (Mer et al., 2000),
homologous recombination (HR) factor Rad52
(Mer et al., 2000), replication checkpoint protein
Tipin (Unsal-Kacmaz et al., 2007), and the
annealing helicase HARP/SMARCAL1 (Bansbach
et al., 2009; Ciccia et al., 2009; Yuan et al., 2009).
These interactions likely aid the multiple roles of
RPA in facilitating DNA repair.
Homology
A close homolog of RPA2, termed RPA4, is located
on Xq21.33 (Haring et al., 2010).
Mutations
Note
Naturally-occurring mutations of human RPA2
have not yet been described. A small number of
genetic polymorphisms have been described in SNP
datasets (Y14S, G15R, and N203S), but these have
not yet been reported to have any biological effects
(NIEHS SNPs program).
RPA2 (replication protein A2, 32kDa) Murphy AKZ, Borowiec JA
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 56
Implicated in
Colorectal adenocarcinoma
Disease
Overexpression of the RPA2 (and RPA1) proteins
have been found to be prognostic indicator of colon
cancer. Strong associations between RPA2
expression and disease stage, lymph node
metastasis, and the histological grade of carcinomas
have been observed.
Prognosis
In addition, RPA2 protein expression correlates
with poor survival of stage II and III patients
(Givalos et al., 2007).
Ductal breast carcinoma
Disease
Levels of anti-RPA2 antibodies was observed to be
significantly higher in sera from breast cancer
patients (10.9%; n = 801) as compared to normal
controls (0.0%; n = 221). Examining individuals
with early stage intraductal in situ carcinomas,
10.3% (n = 39) similarly showed the presence of
high levels of anti-RPA2 antibodies. Even so,
follow-up studies indicated that there were no
apparent differences in mean survival, occurrences
of a second primary tumor, or metastasis frequency
between breast cancer patients that were positive or
negative for anti-RPA2 sera. Although RPA is a
nuclear protein, RPA was seen to be localized to
both nuclei and cytoplasm in the cells of at least
one breast tumor, with RPA also over-expressed
(Tomkiel et al., 2002).
Non-small cell carcinoma
Disease
A fraction of individuals with squamous cell lung
cancer were found to have significant levels of anti-
RPA2 antibodies (9.1%; n = 22) (Tomkiel et al.,
2002).
Laryngeal tumors
Disease
One patient (out of 35; 2.9%) with head and neck
tumors tested positive for the presence of anti-
RPA2 sera (Tomkiel et al., 2002).
Promyelocytic leukemia
Disease
A derivative of the human HL-60 promyelocytic
leukemia cell line (HL-60/P1), selected for its
decreased sensitivity to undergo apoptosis in
response to TNF-related apoptosis-inducing ligand
(TRAIL), was found to have decreased (2-fold)
expression of RPA2 (Petrak et al., 2009).
Sjögren syndrome
Disease
Serum from a patient with Sjögren syndrome was
found to have high levels of anti-RPA2 antibodies.
A higher rate of non-Hodgkin lymphoma, and
lymphoid malignancies, is seen in individuals with
Sjögren syndrome, compared to normal individuals
(Garcia-Lozano et al., 1995).
Systemic lupus erythematosus (SLE)
Disease
One out of 55 individuals with autoimmune
disorders was found to test positive for anti-RPA2
antibodies (1.8%). This individual had SLE, and
secondary Sjögren syndrome (Garcia-Lozano et al.,
1995).
Rheumatoid arthritis (RA)
Note
Fibroblast-like synoviocytes (FLSs) are a cell type
whose invasive properties provide an indicator of
RA severity. Microarray studies from FLSs in DA
rats (arthritis-susceptible inbred model) show a
modest increase in the level of RPA2 mRNA,
compared to back-crossed arthritis-resistant
DA.F344 (Cia5d) congenic strains (Laragione et al.,
2008).
References Kim C, Snyder RO, Wold MS. Binding properties of replication protein A from human and yeast cells. Mol Cell Biol. 1992 Jul;12(7):3050-9
Garcia-Lozano R, Gonzalez-Escribano F, Sanchez-Roman J, Wichmann I, Nuñez-Roldan A. Presence of antibodies to different subunits of replication protein A in autoimmune sera. Proc Natl Acad Sci U S A. 1995 May 23;92(11):5116-20
He Z, Henricksen LA, Wold MS, Ingles CJ. RPA involvement in the damage-recognition and incision steps of nucleotide excision repair. Nature. 1995 Apr 6;374(6522):566-9
Philipova D, Mullen JR, Maniar HS, Lu J, Gu C, Brill SJ. A hierarchy of SSB protomers in replication protein A. Genes Dev. 1996 Sep 1;10(17):2222-33
Bochkarev A, Bochkareva E, Frappier L, Edwards AM. The crystal structure of the complex of replication protein A subunits RPA32 and RPA14 reveals a mechanism for single-stranded DNA binding. EMBO J. 1999 Aug 16;18(16):4498-504
Mer G, Bochkarev A, Gupta R, Bochkareva E, Frappier L, Ingles CJ, Edwards AM, Chazin WJ. Structural basis for the recognition of DNA repair proteins UNG2, XPA, and RAD52 by replication factor RPA. Cell. 2000 Oct 27;103(3):449-56
Bastin-Shanower SA, Brill SJ. Functional analysis of the four DNA binding domains of replication protein A. The role of RPA2 in ssDNA binding. J Biol Chem. 2001 Sep 28;276(39):36446-53
Kalma Y, Marash L, Lamed Y, Ginsberg D. Expression analysis using DNA microarrays demonstrates that E2F-1 up-regulates expression of DNA replication genes including replication protein A2. Oncogene. 2001 Mar 15;20(11):1379-87
Bochkareva E, Korolev S, Lees-Miller SP, Bochkarev A. Structure of the RPA trimerization core and its role in the multistep DNA-binding mechanism of RPA. EMBO J. 2002 Apr 2;21(7):1855-63
RPA2 (replication protein A2, 32kDa) Murphy AKZ, Borowiec JA
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Tomkiel JE, Alansari H, Tang N, Virgin JB, Yang X, VandeVord P, Karvonen RL, Granda JL, Kraut MJ, Ensley JF, Fernández-Madrid F. Autoimmunity to the M(r) 32,000 subunit of replication protein A in breast cancer. Clin Cancer Res. 2002 Mar;8(3):752-8
Binz SK, Sheehan AM, Wold MS. Replication protein A phosphorylation and the cellular response to DNA damage. DNA Repair (Amst). 2004 Aug-Sep;3(8-9):1015-24
Anantha RW, Vassin VM, Borowiec JA. Sequential and synergistic modification of human RPA stimulates chromosomal DNA repair. J Biol Chem. 2007 Dec 7;282(49):35910-23
Givalos N, Gakiopoulou H, Skliri M, Bousboukea K, Konstantinidou AE, Korkolopoulou P, Lelouda M, Kouraklis G, Patsouris E, Karatzas G. Replication protein A is an independent prognostic indicator with potential therapeutic implications in colon cancer. Mod Pathol. 2007 Feb;20(2):159-66
Unsal-Kaçmaz K, Chastain PD, Qu PP, Minoo P, Cordeiro-Stone M, Sancar A, Kaufmann WK. The human Tim/Tipin complex coordinates an Intra-S checkpoint response to UV that slows replication fork displacement. Mol Cell Biol. 2007 Apr;27(8):3131-42
Anantha RW, Sokolova E, Borowiec JA. RPA phosphorylation facilitates mitotic exit in response to mitotic DNA damage. Proc Natl Acad Sci U S A. 2008 Sep 2;105(35):12903-8
Laragione T, Brenner M, Li W, Gulko PS. Cia5d regulates a new fibroblast-like synoviocyte invasion-associated gene expression signature. Arthritis Res Ther. 2008;10(4):R92
Bansbach CE, Bétous R, Lovejoy CA, Glick GG, Cortez D. The annealing helicase SMARCAL1 maintains genome integrity at stalled replication forks. Genes Dev. 2009 Oct 15;23(20):2405-14
Ciccia A, Bredemeyer AL, Sowa ME, Terret ME, Jallepalli PV, Harper JW, Elledge SJ. The SIOD disorder protein
SMARCAL1 is an RPA-interacting protein involved in replication fork restart. Genes Dev. 2009 Oct 15;23(20):2415-25
Feng J, Wakeman T, Yong S, Wu X, Kornbluth S, Wang XF. Protein phosphatase 2A-dependent dephosphorylation of replication protein A is required for the repair of DNA breaks induced by replication stress. Mol Cell Biol. 2009 Nov;29(21):5696-709
Petrak J, Toman O, Simonova T, Halada P, Cmejla R, Klener P, Zivny J. Identification of molecular targets for selective elimination of TRAIL-resistant leukemia cells. From spots to in vitro assays using TOP15 charts. Proteomics. 2009 Nov;9(22):5006-15
Vassin VM, Anantha RW, Sokolova E, Kanner S, Borowiec JA. Human RPA phosphorylation by ATR stimulates DNA synthesis and prevents ssDNA accumulation during DNA-replication stress. J Cell Sci. 2009 Nov 15;122(Pt 22):4070-80
Yuan J, Ghosal G, Chen J. The annealing helicase HARP protects stalled replication forks. Genes Dev. 2009 Oct 15;23(20):2394-9
Haring SJ, Humphreys TD, Wold MS. A naturally occurring human RPA subunit homolog does not support DNA replication or cell-cycle progression. Nucleic Acids Res. 2010 Jan;38(3):846-58
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This article should be referenced as such:
Murphy AKZ, Borowiec JA. RPA2 (replication protein A2, 32kDa). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):54-57.
Gene Section Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 58
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
S100A7 (S100 calcium binding protein A7) Jill I Murray, Martin J Boulanger
Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia
V8W 3P6, Canada (JIM, MJB)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/S100A7ID42194ch1q21.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI S100A7ID42194ch1q21.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity
Other names: PSOR1, S100A7c
HGNC (Hugo): S100A7
Location: 1q21.3
Local order: S100A7 is located on chromosome
1cen-q21 between D1Z5 and MUC1 (Borglum et
al., 1995).
Note: S100A7 is also known as psoriasin, psoriasin
1, S100 calcium binding protein A7, S100-A7,
S100A7c, and PSOR1.
S100A7, a member of the S100 family, was first
identified as a protein upregulated in psoriasis
(Madsen et al., 1991).
DNA/RNA
Note
S100A7 is located on chromosome 1q21 within the
epidermal differentiation complex.
Description
The S100A7 gene has 3 exons and 2 introns with a
genomic structure similar to other S100 family
members. Exon 1 encodes the 5' untranslated region
while exons 2 and 3 contain the protein coding
sequence. Exon 2 encodes the start codon and the
non-canonical N-terminal EF-hand while exon 3
encodes the carboxyl-terminal EF-hand.
Transcription
The S100A7 gene encodes for a single
constitutively spliced transcript. An EST has been
reported in which an alternative promoter is used to
produce an identical S100A7 mRNA (See Ensembl,
UCSC genome browser).
Pseudogene
Five copies of S100A7 in the human genome have
been reported including the closely related paralog
S100A15 (also known as S100A7A) (Kulski et al.,
2003; Wolf et al., 2003). Two of the five reported
copies of S100A7, S100A7d (S100A7P1) and
S100A7e (S100A7P2), are proposed to be non-
coding pseudogenes (Kulski et al., 2003; Marenholz
et al., 2006).
The S100A7 genomic organization includes 3 exons and 2 introns with exons 2 and 3 containing the protein encoding sequence
(Semprini et al., 1999). The EF-hand domains are highlighted (Burgisser et al., 1995).
S100A7 (S100 calcium binding protein A7) Murray JI, Boulanger MJ
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 59
A. S100A7 primary sequence highlighting the calcium- and zinc-binding residues and the EF-hand domains.
B. The 3D structure of zinc- and calcium-bound S100A7 dimer (2psr).
Protein
Note
S100A7 is a member of the S100 family of
calcium-binding signaling proteins. S100A7 has
both intracellular and extracellular functions.
Description
S100A7 is a small 11.4 kDa protein containing a C-
terminal canonical calcium-binding EF-hand motif
and an N-terminal non-canonical EF-hand motif
which is characteristic of the S100 protein family.
S100A7 forms a homodimer with one Ca2+
ion
bound by the canonical EF-hand motif in each
monomer and two Zn2+
ions located at the dimer
interface (Brodersen et al., 1999). S100A7
monomers and putative higher order multimers
have been observed in both psoriatic and healthy
epidermis (Ruse et al., 2001).
Expression
S100A7 is present at low levels in healthy skin,
however it is highly upregulated in psoriatic
epidermal keratinocytes (Madsen et al., 1991). E.
Coli has been shown to induce S100A7 expression
in keratinocytes (Gläser et al., 2005).
S100A7 expression is upregulated in several
cancers including skin, breast, lung, head, neck,
cervix, bladder and gastric cancer (for review see
Emberley et al., 2004).
S100A7 expression is induced in MCF10 cells by
stresses such as serum deprivation and cell
confluency (Enerback et al., 2002).
S100A7 is repressed by BRCA1 in a c-myc
dependent manner in HCC-BR116 cells (Kennedy
et al., 2005). 17beta-estradiol treatment increased
S100A7 expression in an estrogen receptor beta
dependent manner in MCF-7 cells (Skliris et al.,
2007). Epidermal Growth Factor induces S100A7
expression in MCF-7 and MDA-MB-468 cells
(Paruchuri et al., 2008).
S100A7 expression is induced by proinflammatory
cytokines in skin and breast cancer cells. S100A7
expression is enhanced in human keratinocytes by
stimulation with the cytokine IL-22 in combination
with IL-17 or IL-17F (Liang et al., 2006).
Oncostatin-M was shown to induce S100A7
expression in human epidermal cell skin
equivalents (Gazel et al., 2006). S100A7 expression
is induced by the cytokines oncostatin-M and IL-6
in MCF-7, TD47 and MDA-MB-468 cell lines
(West and Watson, 2010).
Localisation
S100A7 is localized to the cytoplasm, nucleus, cell
periphery and is also secreted from cells.
S100A7 (S100 calcium binding protein A7) Murray JI, Boulanger MJ
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 60
In keratinocytes, S100A7 is observed in the
cytoplasm when untreated and at the cell periphery
upon stimulation with calcium (Ruse et al., 2003).
S100A7 is expressed at low levels or is not detected
in healthy breast cells. In breast cancer cells,
however, S100A7 is observed in the nucleus and
cytoplasm and is also secreted (Al-Haddad et al.,
1999; Enerback et al., 2002).
Function
S100A7 has been shown to function as a
chemotactic factor for neutrophils and CD4+ T
cells (Jinquan et al., 1996). S100A7 binds RAGE
(receptor for advanced glycation end products) in a
zinc-dependent manner and is proposed to mediate
chemotaxis in a RAGE-dependent manner (Wolf et
al., 2008). S100A7 present in skin functions as a
Zn-dependent antimicrobial towards E.Coli (Glaser
et al., 2005). S100A7 has also been shown to play
an antibacterial role in wound healing (Lee and
Eckert, 2007). S100A7 is a substrate for
transglutaminase (Ruse et al., 2001).
S100A7 interacts, co-purifies and colocalizes in the
cytoplasm with epidermal-type fatty acid-binding
protein (E-FABP), a protein which is also
upregulated in psoriasis (Hagens et al., 1999; Ruse
et al., 2003). S100A7 has been shown to interact
with RanBPM by yeast two-hybrid and co-
immunoprecipitation studies in breast cancer cells
(Emberley et al., 2002). S100A7 has been shown to
interact with the multifunctional signalling protein,
Jab1, yeast two-hybrid and co-immunoprecipitation
studies in breast cancer cells (Emberley et al.,
2003). The Jab1-S100A7 interaction and
downstream effects were disrupted by mutation of a
Jab1-binding site (Emberley et al., 2003; West et
al., 2009).
Homology
S100A7 is a member of the S100 family of
vertebrate proteins. Among the S100 family,
S100A7 is the most divergent (Burgisser et al.,
1995) with the exception of a recently identified
paralog S100A715 (or S100A7A), with which it
shares 93% similarity (Wolf et al., 2003). A bovine
ortholog to S100A7, Bosd3 (Virtanen, 2006) and
equine ortholog (Leeb et al., 2005) have also been
reported. The mouse S100A7, which has 40%
similarity (Webb et al., 2005), has been assigned
the designation mouse S100A15 (Wolf et al., 2006).
Mutations
Note
An allergy associated polymorphism of S100A7
(rs3014837) has been reported (Bryborn et al.,
2008).
Implicated in
Psoriasis and other skin diseases
Note
S100A7 is associated with inflammation in several
skin diseases (Algermissen et al., 1996). S100A7
was originally identified as a protein secreted from
psoriatic skin (Madsen et al., 1991). S100A7 is also
overexpressed in skin lesions of patients with lichen
sclerosus (Gambichler et al., 2009), acne inversa
(Schlapbach et al., 2009), and middle ear
cholesteatoma (Kim et al., 2009).
Non-melanoma skin cancer
Note
S100A7 may play a role in the progression of skin
cancer. S100A7 expression is not observed in
healthy epidermis. When S100A7 levels were
studied by immunohistochemistry in squamous cell
carcinoma skin lesions, higher levels of expression
were found in pre-invasive squamous cell
carcinoma in situ compared to invasive squamous
cell carcinoma (Alowami et al., 2003). In a separate
study, S100A7 mRNA levels, determined by real-
time PCR, were upregulated in pre-cancerous skin
lesions and epithelial skin tumours including basal
cell carcinoma and squamous cell carcinoma
(Moubayed et al., 2007).
Melanoma
Note
S100A7 protein was observed at higher levels in the
urine of melanoma patients compared to healthy
controls (Brouard et al., 2002), although S100A7
was not detected in melanoma cells (Petersson et
al., 2009).
Ductal carcinoma in situ (DCIS) and breast cancer
Note
S100A7 was first associated with primary breast
cancer (Moog-Lutz et al., 1995). Later studies
identified S100A7 as one of the most highly
expressed genes in DCIS, a key stage before the
transition to invasive breast cancer (Leygue et al.,
1996; Enerback et al., 2002). When S100A7 is
expressed in later stages of breast cancer it is
associated with the agressive estrogen-negative
tumors and poor prognosis (Al-Haddad et al., 1999;
Emberley et al., 2004). In vivo mouse model
studies have shown that S100A7 promotes
tumorigenesis (Emberley et al., 2003; Krop et al.,
2005). Several of the tumorigenic effects of
S100A7, including upregulation of NF-kappaB,
PI3K-Akt, and AP-1 as well as promotion of cell
survival, are mediated by the interaction of S100A7
with Jab1 (Emberley et al., 2003; Emberley et al.,
2005).
S100A7 (S100 calcium binding protein A7) Murray JI, Boulanger MJ
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 61
Epithelial ovarian cancer
Note
S1007 mRNA and protein levels are upregulated in
epithelial ovarian carcinoma tissue compared to
normal and benign ovary tissue (Gagnon et al.,
2008). Autoantibodies to S100A7 were detected at
higher levels in the plasma of early and late-stage
ovarian cancer patients compared to healthy
controls (Gagnon et al., 2008). S100A7
autoantibodies may be useful as a biomarker for
epithelial ovarian cancer (for review see Piura and
Piura, 2009).
Lung squamous cell carcinoma
Note
S100A7 is associated with non-small lung
squamous cell carcinoma metastasis to the brain
(Zhang et al., 2007). Proteomic studies identified
S100A7 as a protein upregulated in a brain
metastasis lung squamous cell carcinoma cell line
and S100A7 overexpression was confirmed in brain
metastasis tissues (Zhang et al., 2007).
Bladder squamous cell carcinoma
Note
S100A7 was detected in bladder squamous cell
carcinoma tumors and also in the urine of patients
with bladder squamous cell carcinoma (Celis et al.,
1996; Ostergaard et al., 1997). As a result, S100A7
has been proposed to be a potential biomarker for
bladder squamous cell carcinoma (Celis et al.,
1996; Ostergaard et al., 1997; Ostergaard et al.,
1999).
Oral squamous cell carcinoma
Note
S100A7 is associated with oral squamous cell
carcinoma (Zhou et al., 2008; Kesting et al., 2009).
RT-PCR and immunofluorescence studies showed
that S100A7 mRNA and protein levels respectively
are up-regulated in oral squamous cell carcinoma
tissues compared to normal oral tissues (Kesting et
al., 2009).
Head-and-neck squamous cell carcinoma
Note
S100A7 is a highly upregulated biomarker in head-
and-neck squamous cell carcinomas (Ralhan et al.,
2008).
Gastric cancer
Note
SAGE (serial analysis of gene expression) studies
identified S100A7 as one of the top twenty genes
upregulated in gastric cancer (El-Rifai et al., 2002).
Further mining of publicly available SAGE, virtual
Northern Blot, and microarray data confirmed the
association of S100 proteins such as S100A7 with
gastric cancer (Liu et al., 2008).
Chronic rhinosinusitis
Note
Chronic rhinosinusitis (CRS) is characterized by a
persistant inflammation of the nasal mucosa. It has
been proposed that the antibacterial function of
S100A7 play a role in protecting against the
environmental factors that contribute to chronic
rinosinusitis (for review see Tieu et al., 2009).
Reduced levels of S100A7 were detected in the
nasal lavage fluid of patients with allergic rhinitis
when compared to controls (Bryborn et al., 2005).
A polymorphism (RS3014837) has been linked
with allergic individuals in Sweden (Bryborn et al.,
2008).
Systemic sclerosis (SSc)
Note
S100A7 is upregulated in the saliva of patients with
systemic sclerosis when compared to healthy
individuals and has been proposed as a potential
biomarker for systemic sclerosis with pulmonary
involvement (Giusti et al., 2007; Baldini et al.,
2008).
Alzheimer's disease
Note
A recent study has suggested that S100A7 is a
potential biomarker for Alzheimer's disease.
Increased levels of S100A7 were detected in the
cerebralspinal fluid and brain of patients with
Alzheimer's disease (Qin et al., 2009).
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Børglum AD, Flint T, Madsen P, Celis JE, Kruse TA. Refined mapping of the psoriasin gene S100A7 to chromosome 1cen-q21. Hum Genet. 1995 Nov;96(5):592-6
Bürgisser DM, Siegenthaler G, Kuster T, Hellman U, Hunziker P, Birchler N, Heizmann CW. Amino acid sequence analysis of human S100A7 (psoriasin) by tandem mass spectrometry. Biochem Biophys Res Commun. 1995 Dec 5;217(1):257-63
Moog-Lutz C, Bouillet P, Régnier CH, Tomasetto C, Mattei MG, Chenard MP, Anglard P, Rio MC, Basset P. Comparative expression of the psoriasin (S100A7) and S100C genes in breast carcinoma and co-localization to human chromosome 1q21-q22. Int J Cancer. 1995 Oct 9;63(2):297-303
Algermissen B, Sitzmann J, LeMotte P, Czarnetzki B. Differential expression of CRABP II, psoriasin and cytokeratin 1 mRNA in human skin diseases. Arch Dermatol Res. 1996 Jul;288(8):426-30
Celis JE, Rasmussen HH, Vorum H, Madsen P, Honoré B, Wolf H, Orntoft TF. Bladder squamous cell carcinomas express psoriasin and externalize it to the urine. J Urol. 1996 Jun;155(6):2105-12
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Leygue E, Snell L, Hiller T, Dotzlaw H, Hole K, Murphy LC, Watson PH. Differential expression of psoriasin messenger RNA between in situ and invasive human breast carcinoma. Cancer Res. 1996 Oct 15;56(20):4606-9
Ostergaard M, Rasmussen HH, Nielsen HV, Vorum H, Orntoft TF, Wolf H, Celis JE. Proteome profiling of bladder squamous cell carcinomas: identification of markers that define their degree of differentiation. Cancer Res. 1997 Sep 15;57(18):4111-7
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Brodersen DE, Nyborg J, Kjeldgaard M. Zinc-binding site of an S100 protein revealed. Two crystal structures of Ca2+-bound human psoriasin (S100A7) in the Zn2+-loaded and Zn2+-free states. Biochemistry. 1999 Feb 9;38(6):1695-704
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Gläser R, Harder J, Lange H, Bartels J, Christophers E, Schröder JM. Antimicrobial psoriasin (S100A7) protects human skin from Escherichia coli infection. Nat Immunol. 2005 Jan;6(1):57-64
Kennedy RD, Gorski JJ, Quinn JE, Stewart GE, James CR, Moore S, Mulligan K, Emberley ED, Lioe TF, Morrison PJ, Mullan PB, Reid G, Johnston PG, Watson PH, Harkin DP. BRCA1 and c-Myc associate to transcriptionally repress psoriasin, a DNA damage-inducible gene. Cancer Res. 2005 Nov 15;65(22):10265-72
Krop I, März A, Carlsson H, Li X, Bloushtain-Qimron N, Hu M, Gelman R, Sabel MS, Schnitt S, Ramaswamy S, Kleer CG, Enerbäck C, Polyak K. A putative role for psoriasin in breast tumor progression. Cancer Res. 2005 Dec 15;65(24):11326-34
Leeb T, Bruhn O, Philipp U, Kuiper H, Regenhard P, Paul S, Distl O, Chowdhary BP, Kalm E, Looft C. Assignment of the equine S100A7 gene (psoriasin 1) to chromosome 5p12-->p13 by fluorescence in situ hybridization and radiation hybrid mapping. Cytogenet Genome Res. 2005;109(4):533
Webb M, Emberley ED, Lizardo M, Alowami S, Qing G, Alfia'ar A, Snell-Curtis LJ, Niu Y, Civetta A, Myal Y, Shiu R, Murphy LC, Watson PH. Expression analysis of the mouse S100A7/psoriasin gene in skin inflammation and mammary tumorigenesis. BMC Cancer. 2005 Feb 17;5:17
Gazel A, Rosdy M, Bertino B, Tornier C, Sahuc F, Blumenberg M. A characteristic subset of psoriasis-associated genes is induced by oncostatin-M in reconstituted epidermis. J Invest Dermatol. 2006 Dec;126(12):2647-57
Liang SC, Tan XY, Luxenberg DP, Karim R, Dunussi-Joannopoulos K, Collins M, Fouser LA. Interleukin (IL)-22 and IL-17 are coexpressed by Th17 cells and cooperatively enhance expression of antimicrobial peptides. J Exp Med. 2006 Oct 2;203(10):2271-9
Marenholz I, Lovering RC, Heizmann CW. An update of the S100 nomenclature. Biochim Biophys Acta. 2006 Nov;1763(11):1282-3
S100A7 (S100 calcium binding protein A7) Murray JI, Boulanger MJ
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 63
Virtanen T. Psoriasin and its allergenic bovine homolog Bos d 3. Cell Mol Life Sci. 2006 May;63(10):1091-4
Wolf R, Voscopoulos CJ, FitzGerald PC, Goldsmith P, Cataisson C, Gunsior M, Walz M, Ruzicka T, Yuspa SH. The mouse S100A15 ortholog parallels genomic organization, structure, gene expression, and protein-processing pattern of the human S100A7/A15 subfamily during epidermal maturation. J Invest Dermatol. 2006 Jul;126(7):1600-8
Giusti L, Bazzichi L, Baldini C, Ciregia F, Mascia G, Giannaccini G, Del Rosso M, Bombardieri S, Lucacchini A. Specific proteins identified in whole saliva from patients with diffuse systemic sclerosis. J Rheumatol. 2007 Oct;34(10):2063-9
Lee KC, Eckert RL. S100A7 (Psoriasin)--mechanism of antibacterial action in wounds. J Invest Dermatol. 2007 Apr;127(4):945-57
Moubayed N, Weichenthal M, Harder J, Wandel E, Sticherling M, Gläser R. Psoriasin (S100A7) is significantly up-regulated in human epithelial skin tumours. J Cancer Res Clin Oncol. 2007 Apr;133(4):253-61
Skliris GP, Lewis A, Emberley E, Peng B, Weebadda WK, Kemp A, Davie JR, Shiu RP, Watson PH, Murphy LC. Estrogen receptor-beta regulates psoriasin (S100A7) in human breast cancer. Breast Cancer Res Treat. 2007 Jul;104(1):75-85
Zhang H, Wang Y, Chen Y, Sun S, Li N, Lv D, Liu C, Huang L, He D, Xiao X. Identification and validation of S100A7 associated with lung squamous cell carcinoma metastasis to brain. Lung Cancer. 2007 Jul;57(1):37-45
Baldini C, Giusti L, Bazzichi L, Ciregia F, Giannaccini G, Giacomelli C, Doveri M, Del Rosso M, Bombardieri S, Lucacchini A. Association of psoriasin (S100A7) with clinical manifestations of systemic sclerosis: is its presence in whole saliva a potential predictor of pulmonary involvement? J Rheumatol. 2008 Sep;35(9):1820-4
Bryborn M, Halldén C, Säll T, Adner M, Cardell LO. Comprehensive evaluation of genetic variation in S100A7 suggests an association with the occurrence of allergic rhinitis. Respir Res. 2008 Mar 28;9:29
Gagnon A, Kim JH, Schorge JO, Ye B, Liu B, Hasselblatt K, Welch WR, Bandera CA, Mok SC. Use of a combination of approaches to identify and validate relevant tumor-associated antigens and their corresponding autoantibodies in ovarian cancer patients. Clin Cancer Res. 2008 Feb 1;14(3):764-71
Liu J, Li X, Dong GL, Zhang HW, Chen DL, Du JJ, Zheng JY, Li JP, Wang WZ. In silico analysis and verification of S100 gene expression in gastric cancer. BMC Cancer. 2008 Sep 16;8:261
Paruchuri V, Prasad A, McHugh K, Bhat HK, Polyak K, Ganju RK. S100A7-downregulation inhibits epidermal growth factor-induced signaling in breast cancer cells and blocks osteoclast formation. PLoS One. 2008 Mar 5;3(3):e1741
Ralhan R, Desouza LV, Matta A, Chandra Tripathi S, Ghanny S, Datta Gupta S, Bahadur S, Siu KW. Discovery and verification of head-and-neck cancer biomarkers by differential protein expression analysis using iTRAQ labeling, multidimensional liquid chromatography, and tandem mass spectrometry. Mol Cell Proteomics. 2008 Jun;7(6):1162-73
Wolf R, Howard OM, Dong HF, Voscopoulos C, Boeshans K, Winston J, Divi R, Gunsior M, Goldsmith P, Ahvazi B, Chavakis T, Oppenheim JJ, Yuspa SH. Chemotactic activity of S100A7 (Psoriasin) is mediated by the receptor for advanced glycation end products and potentiates inflammation with highly homologous but functionally distinct S100A15. J Immunol. 2008 Jul 15;181(2):1499-506
Zhou G, Xie TX, Zhao M, Jasser SA, Younes MN, Sano D, Lin J, Kupferman ME, Santillan AA, Patel V, Gutkind JS, Ei-Naggar AK, Emberley ED, Watson PH, Matsuzawa SI, Reed JC, Myers JN. Reciprocal negative regulation between S100A7/psoriasin and beta-catenin signaling plays an important role in tumor progression of squamous cell carcinoma of oral cavity. Oncogene. 2008 Jun 5;27(25):3527-38
Gambichler T, Skrygan M, Tigges C, Kobus S, Gläser R, Kreuter A. Significant upregulation of antimicrobial peptides and proteins in lichen sclerosus. Br J Dermatol. 2009 Nov;161(5):1136-42
Kesting MR, Sudhoff H, Hasler RJ, Nieberler M, Pautke C, Wolff KD, Wagenpfeil S, Al-Benna S, Jacobsen F, Steinstraesser L. Psoriasin (S100A7) up-regulation in oral squamous cell carcinoma and its relation to clinicopathologic features. Oral Oncol. 2009 Aug;45(8):731-6
Kim KH, Cho JG, Song JJ, Woo JS, Lee HM, Jung HH, Hwang SJ, Chae S. Psoriasin (S100A7), an antimicrobial peptide, is increased in human middle ear cholesteatoma. Acta Otolaryngol. 2009 Aug 25;:1-5
Petersson S, Shubbar E, Enerbäck L, Enerbäck C. Expression patterns of S100 proteins in melanocytes and melanocytic lesions. Melanoma Res. 2009 Aug;19(4):215-25
Piura B, Piura E. Autoantibodies to tumor-associated antigens in epithelial ovarian carcinoma. J Oncol. 2009;2009:581939
Qin W, Ho L, Wang J, Peskind E, Pasinetti GM. S100A7, a novel Alzheimer's disease biomarker with non-amyloidogenic alpha-secretase activity acts via selective promotion of ADAM-10. PLoS One. 2009;4(1):e4183
Schlapbach C, Yawalkar N, Hunger RE. Human beta-defensin-2 and psoriasin are overexpressed in lesions of acne inversa. J Am Acad Dermatol. 2009 Jul;61(1):58-65
Tieu DD, Kern RC, Schleimer RP. Alterations in epithelial barrier function and host defense responses in chronic rhinosinusitis. J Allergy Clin Immunol. 2009 Jul;124(1):37-42
West NR, Farnell B, Murray JI, Hof F, Watson PH, Boulanger MJ. Structural and functional characterization of a triple mutant form of S100A7 defective for Jab1 binding. Protein Sci. 2009 Dec;18(12):2615-23
West NR, Watson PH. S100A7 (psoriasin) is induced by the proinflammatory cytokines oncostatin-M and interleukin-6 in human breast cancer. Oncogene. 2010 Apr 8;29(14):2083-92
This article should be referenced as such:
Murray JI, Boulanger MJ. S100A7 (S100 calcium binding protein A7). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):58-63.
Gene Section Mini Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 64
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
SOX10 (SRY (sex determining region Y)-box 10) Michael Wegner
Institut fuer Biochemie, Emil-Fischer-Zentrum, Universitaet Erlangen-Nuernberg, 91054 Erlangen,
Germany (MW)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/SOX10ID43768ch22q13.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI SOX10ID43768ch22q13.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity
Other names: DOM, MGC15649, WS2E, WS4
HGNC (Hugo): SOX10
Location: 22q13.1
Local order: Flanked by POLR2F (DNA-directed
RNA polymerase II polypeptide F) and PICK1
(protein interacting with PRKCA 1).
DNA/RNA
Note
SOX10 was first identified as a gene mutated in
patients suffering from Waardenburg syndrome
type 4C (WS4C). SOX10 mutations also cause
Waardenburg syndrome type 2E (WS2E) with or
without neurologic involvement, Yemenite deaf-
blind hypopigmentation syndrome and PCWH
syndrome. They usually occur in the heterozygous
state and can be either sporadic or familial.
Description
DNA size: 12.22kb; 5 Exons.
Transcription
mRNA size: 2882 nucleotides.
Protein
Note
The SOX10 protein belongs to subgroup E of the
SOX protein family. All 20 human members of this
protein family possess a high-mobility-group
(HMG) domain with three alpha-helical regions and
close similarity to the one found in the male sex
determining factor SRY. SOX10 functions as
transcription factor and structural protein in
chromatin. SOX9 and SOX8 are its closest relatives
among human SOX proteins.
Description
SOX10 consists of 466 amino acids. The following
domains exist (from amino terminal to carboxy
terminal): DNA-dependent dimerization domain
(amino acids 61-101), DNA-binding HMG-domain
(amino acids 101-180), context-dependent
transactivation domain K2 (amino acids 233-306)
and main transactivation domain TA (amino acids
400-462). SOX10 possesses two nuclear
localization signals (NLS) at the beginning and the
end of the HMG domain and a nuclear export
sequence (NES in the middle).
The SOX10 gene with its 5 exons. The open reading frame (orange) is split between exons 3-5. The 5' untranslated region is
generated from exons 1-3 and the 3' untranslated region corresponds to the hind part of exon 5.
SOX10 (SRY (sex determining region Y)-box 10) Wegner M
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 65
Human SOX10 and its domains including the DNA-dependent dimerization domain (Dim), the DNA-binding HMG domain, the
context-dependent transactivation domain K2 and the main transactivation domain (TA). Numbers indicate amino acid positions. The bottom shows the exact amino acid sequence of the HMG domain with its three alpha-helices, the 2 nuclear localization
signals (NLS1, NLS2) and the nuclear export sequence (NES).
Expression
SOX10 expression is first detected during
embryonic development in the emerging neural
crest and continues transiently or permanently in
many non-ectomesenchymal derivates of the neural
crest including melanocytes, adrenal medulla and
the developing peripheral nervous system. Within
the developing central nervous system SOX10
marks cells of the oligodendrocyte lineage. In the
adult, SOX10 is predominantly found in
oligodendrocytes, peripheral glial cells,
melanocytes and adult neural crest stem cell
populations.
Localisation
SOX10 is predominantly found in the nucleus as
expected for a transcription factor but possesses the
ability to shuttle between cytoplasm and nucleus
because of the presence of both NLS and NES in
the protein.
Function
SOX10 has multiple roles during development. In
neural crest stem cells, SOX10 is needed for self-
renewal, survival and maintenance of pluripotency.
SOX10 is furthermore required for specification of
melanocytes and peripheral glia from the neural
crest. After specification, SOX10 continues to be
essential for lineage progression and maintenance
of identity in peripheral glia. Terminal
differentiation of oligodendrocytes also depends on
SOX10. SOX10 exerts these functions through
interactions with different sets of transcription
factors. SOX10 probably shares further roles with
its close relatives SOX9 and SOX8 with which it is
co-expressed in several cell types and functions in a
partly redundant manner.
Homology
SOX10 is highly conserved among vertebrates.
Human SOX10 shares 98% identity with Mus
musculus Sox10, 97% identity with Sox10 from
Rattus norvegicus and Canis lupus familiaris, 96%
identity with Bos taurus Sox10 and 82% identity
with Gallus gallus Sox10.
Mutations
Note
SOX10 mutations have so far primarily been
identified as a cause for neurocristopathies
including WS4C, WS2E with or without neurologic
involvement, PCWH syndrome and Yemenite deaf-
bling hypopigmentation syndrome.
Germinal
Missense mutations: S135T, A157V, Q174P.
Nonsense mutations: R43X, T83X, T173X,
E189X, T207X, Q234X, Q250X, S251X, T313X,
S346X, Q364X, Q372X, S376X, Q377X.
Insertions: (L160 R161) dup.
Carboxy terminal extensions: X467C ext82,
X467L ext86, X467T ext86.
Frameshift mutations: S17C fsX17, E57S fsX57,
A110L fsX2, P169R fsX117, R215P fsX64, R261A
fsX25, G266A fsX20, I271S fsX15, H283L fsX11,
H306T fsX5, G308A fsX3, V350C fsX52, A354P
fsX3, E359D fsX42, Q399V fsX2.
Splice mutations: int3 pos.428 +2T>G, int4
pos.698 -2A>C.
Somatic
Missense mutations: R43Q, Q125X, A361V,
G413S, G413D, H414Y, A424V.
Frameshift mutations: P14P fsX10, S449S fsX66.
Implicated in
Melanoma
Note
SOX10 is expressed homogenously in primary and
metastatic melanoma and was identified as a
melanoma tumor antigen. It is often co-expressed
with its relative SOX9. Somatic SOX10 mutations
occur in early stage melanoma. SOX10 upregulates
MITF, MET and Nestin expression in melanoma
and responds to Wnt signals. Its nuclear localization
is controlled by the Tam tyrosine receptor kinase
Tyro3 and its activity is modulated by the
transcription factor SOX5. In sentinel lymph nodes,
SOX10 is a reliable marker for metastatic
melanoma.
SOX10 (SRY (sex determining region Y)-box 10) Wegner M
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 66
Clear cell sarcoma
Note
SOX10 is widely expressed in clear cell sarcoma
where it cooperates with EWS-ATF1 fusions in
MITF activation.
Malignant nerve sheath tumor (MNST)
Note
SOX10 is present in MNST. Expression levels
appear lower than the ones in plexiform
neurofibromas from which MNST arise or in
Schwann cells. SOX10 expression levels are
positively correlated with ErbB3 levels and
inversely correlated with SOX9 levels.
Schwannoma
Note
Homogenous SOX10 expression has been detected
throughout this neoplasm.
Ganglioneuroma
Note
SOX10 expression has been detected. Levels
decrease with increasing grade.
Glioma
Note
SOX10 transcripts and protein were found in
astrocytoma, oligodendroglioma and glioblastoma.
Expression levels and number of expressing cells
within the tumor usually diminish with advancing
grade and malignant progression. SOX10 levels are
particularly high in pilocytic astrocytoma. No
correlation with 1p and 19q deletions has been
detected. In a mouse model, SOX10 has been found
to act synergistically with PDGF during glioma
development, although it was not sufficient to
induce gliomagenesis on its own.
References Pingault V, Bondurand N, Kuhlbrodt K, Goerich DE, Préhu MO, Puliti A, Herbarth B, Hermans-Borgmeyer I, Legius E, Matthijs G, Amiel J, Lyonnet S, Ceccherini I, Romeo G, Smith JC, Read AP, Wegner M, Goossens M. SOX10 mutations in patients with Waardenburg-Hirschsprung disease. Nat Genet. 1998 Feb;18(2):171-3
Khong HT, Rosenberg SA. The Waardenburg syndrome type 4 gene, SOX10, is a novel tumor-associated antigen identified in a patient with a dramatic response to immunotherapy. Cancer Res. 2002 Jun 1;62(11):3020-3
Mollaaghababa R, Pavan WJ. The importance of having your SOX on: role of SOX10 in the development of neural crest-derived melanocytes and glia. Oncogene. 2003 May 19;22(20):3024-34
Inoue K, Khajavi M, Ohyama T, Hirabayashi S, Wilson J, Reggin JD, Mancias P, Butler IJ, Wilkinson MF, Wegner M,
Lupski JR. Molecular mechanism for distinct neurological phenotypes conveyed by allelic truncating mutations. Nat Genet. 2004 Apr;36(4):361-9
Gershon TR, Oppenheimer O, Chin SS, Gerald WL. Temporally regulated neural crest transcription factors distinguish neuroectodermal tumors of varying malignancy and differentiation. Neoplasia. 2005 Jun;7(6):575-84
Wegner M, Stolt CC. From stem cells to neurons and glia: a Soxist's view of neural development. Trends Neurosci. 2005 Nov;28(11):583-8
Addo-Yobo SO, Straessle J, Anwar A, Donson AM, Kleinschmidt-Demasters BK, Foreman NK. Paired overexpression of ErbB3 and Sox10 in pilocytic astrocytoma. J Neuropathol Exp Neurol. 2006 Aug;65(8):769-75
Bannykh SI, Stolt CC, Kim J, Perry A, Wegner M. Oligodendroglial-specific transcriptional factor SOX10 is ubiquitously expressed in human gliomas. J Neurooncol. 2006 Jan;76(2):115-27
Davis IJ, Kim JJ, Ozsolak F, Widlund HR, Rozenblatt-Rosen O, Granter SR, Du J, Fletcher JA, Denny CT, Lessnick SL, Linehan WM, Kung AL, Fisher DE. Oncogenic MITF dysregulation in clear cell sarcoma: defining the MiT family of human cancers. Cancer Cell. 2006 Jun;9(6):473-84
Kelsh RN. Sorting out Sox10 functions in neural crest development. Bioessays. 2006 Aug;28(8):788-98
Ferletta M, Uhrbom L, Olofsson T, Pontén F, Westermark B. Sox10 has a broad expression pattern in gliomas and enhances platelet-derived growth factor-B--induced gliomagenesis. Mol Cancer Res. 2007 Sep;5(9):891-7
Nonaka D, Chiriboga L, Rubin BP. Sox10: a pan-schwannian and melanocytic marker. Am J Surg Pathol. 2008 Sep;32(9):1291-8
Blochin E, Nonaka D. Diagnostic value of Sox10 immunohistochemical staining for the detection of metastatic melanoma in sentinel lymph nodes. Histopathology. 2009 Nov;55(5):626-8
Cronin JC, Wunderlich J, Loftus SK, Prickett TD, Wei X, Ridd K, Vemula S, Burrell AS, Agrawal NS, Lin JC, Banister CE, Buckhaults P, Rosenberg SA, Bastian BC, Pavan WJ, Samuels Y. Frequent mutations in the MITF pathway in melanoma. Pigment Cell Melanoma Res. 2009 Aug;22(4):435-44
Zhu S, Wurdak H, Wang Y, Galkin A, Tao H, Li J, Lyssiotis CA, Yan F, Tu BP, Miraglia L, Walker J, Sun F, Orth A, Schultz PG, Wu X. A genomic screen identifies TYRO3 as a MITF regulator in melanoma. Proc Natl Acad Sci U S A. 2009 Oct 6;106(40):17025-30
Mascarenhas JB, Littlejohn EL, Wolsky RJ, Young KP, Nelson M, Salgia R, Lang D. PAX3 and SOX10 activate MET receptor expression in melanoma. Pigment Cell Melanoma Res. 2010 Apr;23(2):225-37
This article should be referenced as such:
Wegner M. SOX10 (SRY (sex determining region Y)-box 10). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):64-66.
Gene Section Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 67
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
TNFSF18 (tumor necrosis factor (ligand) superfamily, member 18) Theresa Placke, Hans-Georg Kopp, Benjamin Joachim Schmiedel, Helmut Rainer Salih
Eberhard Karls University of Tuebingen, Department of Hematology/Oncology, Otfried-Mueller-Str.
10, 72076 Tuebingen, Germany (TP, HGK, BJS, HRS)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/TNFSF18ID42639ch1q25.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI TNFSF18ID42639ch1q25.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity
Other names: AITRL, GITRL, MGC138237, TL6
HGNC (Hugo): TNFSF18
Location: 1q25.1
DNA/RNA
Description
2 transcript versions published:
mRNA 748 bp: 3 exons (1-223, 224-254, 255-748)
-> coding for 199 aa (2-601),
mRNA 610 bp: 3 exons (1-176, 177-207, 208-610)
-> coding for 177 aa (21-554).
Transcription
Accurate start codon is not clearly defined, 2
transcript versions are published (differing start
codons in exon 1).
Pseudogene
Unknown.
Protein
Description
TNFSF18/GITR ligand (GITRL) is a single-pass
type II transmembrane protein and contains 2
potential glycosylation sites (predicted at 129 aa
and 161 aa). TNFSF18 encompasses 177 or 199 aa
and thus has a molecular weight of about 20 kDa.
In the 177 aa long version, amino acids 1-28
constitute the cytoplasmic domain, 29-49 the
transmembrane domain, and 50-177 the
extracellular domain, whereas in the 199 aa long
variant the amino acids 1-50 constitute the
cytoplasmic domain, 51-71 the transmembrane
domain, and 72-199 the extracellular domain.
Expression
TNFSF18 is expressed on DC, monocytes,
macrophages, B cells, activated T cells, endothelial
cells, osteoclasts and various healthy non-lymphoid
tissues (e.g., testis, ...).
Figure 1. Schematic illustration of the gene structure of human TNFSF18 on chromosome 1. Both published transcript variants are shown. Red boxes represent the mRNA transcript within the gene. The smaller boxes at the beginning and the end of the
transcripts indicate untranslated regions, while the larger boxes display the translated parts.
TNFSF18 (tumor necrosis factor (ligand) superfamily, member 18)
Placke T, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 68
Figure 2. Schematic illustration of the structure of TNFSF18 protein versions, according to the two published transcripts.
The fact that TNFSF18 is constitutively expressed
on resting antigen-presenting cells distinguishes it
from most other TNF family members, which are
not detectable in resting state and are upregulated
following activation.
In addition, TNFSF18 is constitutively expressed
and released as soluble form by solid tumors of
different histological origin and various
hematopoietic malignancies.
Localisation
TNFSF18 is a type II transmembrane protein. A
soluble form of the molecule has been shown to be
released by a yet unknown mechanism e.g. by
tumor cells.
Function
TNFSF18 is the only known ligand for GITR
(TNFRSF18, AITR), which is mainly expressed by
lymphatic cells like T lymphocytes and NK cells.
Upon interaction with its receptor, TNFSF18 is,
like many other TNF family members, capable to
transduce bidirectional signals, i.e. in the receptor
and the ligand bearing cell. Transduction of signals
into TNFSF18 bearing cells has been shown to
cause differentiation of osteoclasts, to activate
macrophages and to alter cytokine production of
healthy myeloid cells, but also of carcinoma and
leukemia cells and influences apoptosis. Activation
of macrophages via TNFSF18 results in increased
secretion of inflammatory mediators like MMP-9,
NO and TNF. In healthy macrophages and myeloid
leukemia cells, TNFSF18 signaling has been found
to involve the MAP kinase pathway.
Binding to TNFRSF18 may induce signaling
through this receptor, which, in mice, has been
implicated in the development of autoimmune
diseases, graft versus host disease and in the
immune response against infectious pathogens and
tumors.
Available data suggest that TNFRSF18 may
mediate different effects in mice and men, and most
functional studies regarding the role of TNFRSF18
in tumor immunology have been performed using
agonistic antibodies or injection of adenovirus
expressing recombinant TNFSF18 into tumors,
which might not reflect the consequences of
TNFRSF18 interaction with its natural ligand in
vivo. In line, studies evaluating immune responses
in GITR-/-
mice have so far not led to a clear picture
of the role of TNFRSF18 in normal physiology.
Homology
The TNFSF18 gene is conserved in human,
chimpanzee, dog, mouse, and rat. The homology
among the other TNF family members is highest
with OX40L.
Mutations
Note
No published single nucleotide polymorphisms
(SNPs).
Implicated in
Host-tumor interaction
Note
In mice, it has been shown that application of the
agonistic GITR antibody DTA-1 delays tumor
progression and can even lead to complete tumor
rejection. Similar results were obtained by using
GITRL-Fc fusion protein. Transfection of tumor
cells with GITRL causes rejection of the tumor and
prolonged survival, while parental cells cannot be
rejected. This effect can be reversed by
administration of a blocking GITRL antibody.
There is evidence that expression of GITRL
promotes the development of tumor-specific T
cells. Re-challenge of mice which once successfully
rejected GITRL-positive tumor results in complete
rejection of both transfected and non-transfected
tumors. Several studies showed increased
infiltration of CD8+ cells in GITRL-expressing
tumors. By the use of depletion experiments and
athymic nude mice it has been shown that for
GITR-GITRL dependent rejection of tumors both
CD4+ and CD8+ T cells as well as NK cells are
required.
TNFSF18 (tumor necrosis factor (ligand) superfamily, member 18)
Placke T, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 69
In humans, controversial data regarding the
function of GITR and GITRL in tumor
immunology were described. Hanabuchi et al.
reported that NK cells are activated by engagement
of GITRL on plasmacytoid dendritic cells, which
can be blocked by anti-GITRL antibody. In
contrast, Baltz et al. and Baessler et al.
demonstrated substantial GITRL expression on
tumor cells and leukemic blasts resulting in
diminished NK cell reactivity. Blockade of GITR-
GITRL interaction by anti-GITR antibody
abrogated the inhibitory effect of GITRL.
Furthermore, stimulation of GITRL substantially
induced the production of TGF-beta and IL-10 by
tumor cells and leukemic blasts. Additionally, they
reported that human GITRL is released by tumor
cells in a soluble form which impairs NK cell
reactivity alike the membrane-bound form. Thus,
GITRL expression seems to affect the interaction of
human tumor cells with the immune system by
influencing tumor cell immunogenity and
metastasis and creating an immunosuppressive
cytokine microenvironment. The inhibitory effect
of GITRL on human NK cells was further
supported by Liu et al., who reported inhibition of
NK cell proliferation and cytokine production and
increased apoptosis after GITR stimulation. These
controversial data regarding the function of GITR
on human NK cells may be due to the usage of
different reagents and different experimental
condition.
The results regarding the role of GITR and GITRL
in tumor immunology are controversial in mice and
humans. Thus, GITR and GITRL may mediate
different effects in mice and men, and in line
suppression of human regulatory T cells, in contrast
to their murine counterparts, is not inhibited by
GITR. Many studies employed agonistic antibodies
or recombinant protein for GITR stimulation and
not constitutively GITRL-expressing cells. Thus,
these studies do not involve possible influences of
reverse signaling mediated by GITRL, which may
change reaction of GITRL-bearing cells and may in
turn alter functions of GITR-bearing cells.
Autoimmune disease
Note
The influence of GITR and GITRL was tested in
different mouse models of autoimmune disease.
Onset of autoimmune diabetes in NOD mice is
accelerated if they are treated with agonistic GITR
mAb, and activation of CD4+ T cells is increased
compared to control treated mice. Likewise,
application of a blocking GITRL antibody protected
from diabetes. In GITR -/- mice, experimental
autoimmune diseases take an attenuated course.
GITR -/- mice with collagen-induced arthritis show
less joint inflammation and bone erosion than
wildtype mice. Furthermore, lower concentrations
of inflammatory mediators were reported. In line
with these findings, GITR triggering antibody
exacerbates collagen-induced arthritis in wildtype
mice compared to control-treated siblings.
However, all these studies regarding the function of
GITR and its ligand in autoimmune disease were
performed in mice. Further investigation is needed
to elucidate the relevance of GITR and GITR ligand
in human autoimmune disease and to clarify the
similarities and differences of these molecules in
mice and men.
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Shin HH, Lee MH, Kim SG, Lee YH, Kwon BS, Choi HS. Recombinant glucocorticoid induced tumor necrosis factor receptor (rGITR) induces NOS in murine macrophage. FEBS Lett. 2002 Mar 13;514(2-3):275-80
Spinicelli S, Nocentini G, Ronchetti S, Krausz LT, Bianchini R, Riccardi C. GITR interacts with the pro-apoptotic protein Siva and induces apoptosis. Cell Death Differ. 2002 Dec;9(12):1382-4
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Baumgartner-Nielsen J, Vestergaard C, Thestrup-Pedersen K, Deleuran M, Deleuran B. Glucocorticoid-induced tumour necrosis factor receptor (GITR) and its ligand (GITRL) in atopic dermatitis. Acta Derm Venereol. 2006;86(5):393-8
Cohen AD, Diab A, Perales MA, Wolchok JD, Rizzuto G, Merghoub T, Huggins D, Liu C, Turk MJ, Restifo NP, Sakaguchi S, Houghton AN. Agonist anti-GITR antibody enhances vaccine-induced CD8(+) T-cell responses and tumor immunity. Cancer Res. 2006 May 1;66(9):4904-12
Cuzzocrea S, Nocentini G, Di Paola R, Agostini M, Mazzon E, Ronchetti S, Crisafulli C, Esposito E, Caputi AP, Riccardi C. Proinflammatory role of glucocorticoid-induced TNF receptor-related gene in acute lung inflammation. J Immunol. 2006 Jul 1;177(1):631-41
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Ramirez-Montagut T, Chow A, Hirschhorn-Cymerman D, Terwey TH, Kochman AA, Lu S, Miles RC, Sakaguchi S, Houghton AN, van den Brink MR. Glucocorticoid-induced TNF receptor family related gene activation overcomes tolerance/ignorance to melanoma differentiation antigens and enhances antitumor immunity. J Immunol. 2006 Jun 1;176(11):6434-42
Baltz KM, Krusch M, Bringmann A, Brossart P, Mayer F, Kloss M, Baessler T, Kumbier I, Peterfi A, Kupka S, Kroeber S, Menzel D, Radsak MP, Rammensee HG, Salih HR. Cancer immunoediting by GITR (glucocorticoid-induced TNF-related protein) ligand in humans: NK cell/tumor cell interactions. FASEB J. 2007 Aug;21(10):2442-54
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This article should be referenced as such:
Placke T, Kopp HG, Schmiedel BJ, Salih HR. TNFSF18 (tumor necrosis factor (ligand) superfamily, member 18). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):67-71.
Gene Section Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 72
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
USF1 (upstream transcription factor 1) Adrie JM Verhoeven
Cardiovascular Research School (COEUR), Department of Biochemistry, Erasmus MC, Rotterdam,
Netherlands (AJMV)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/USF1ID45856ch1q23.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI USF1ID45856ch1q23.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity Other names: bHLHb11, FCHL, FCHL1,
HYPLIP1, MLTF, MLTF1, UEF
HGNC (Hugo): USF1
Location: 1q23.3
Local order:
From centromere to telomere:
F11R (F11 receptor) (on reverse strand), TSTD1
(thiosulfate sulfurtransferase (rhodanese)-like
domain containing 1) (on reverse strand), USF1
(upstream transcription factor 1) (on reverse
strand), ARHGAP30 (Rho GTPase activating
protein 30) (on reverse strand), PVRL4 (poliovirus
receptor-related 4) (on reverse strand), KLHDC9
(kelch domain containing 9) (on plus strand),
PFDN2 (prefoldin subunit 2) (on reverse strand).
Note
USF1 is a bHLH-ZIP transcription factor which
forms homo-dimers or heterodimers with USF2, a
highly homologous bHLH-ZIP transcription factor.
USF1 and USF2 homo- and heterodimers are
similarly active in affecting transcription of most
target genes. USF2 homodimers may have
additional effects.
DNA/RNA
Description
The human USF1 gene on chromosome 1q23 spans
6.73 kb and 11 exons.
Transcription
The mRNA is about 1870 nt. Translation is from a
start codon in exon 2 and ends at a stop codon in
exon 11, and results in a 310 amino acid protein
product. In a splice variant, an alternative donor
splice site within exon 4 is used; translation from
this variant mRNA is from an in-frame start codon
in exon 5, and results in a 251 amino acid protein
product (Saito et al., 2003).
Human USF1 gene diagram. Exons 1 through 11 are depicted by boxes, the open reading frames of the USF1 protein and the
splice variant are shown by dark and light green colour code, respectively. The approximate positions of two functional SNPs are also indicated.
USF1 (upstream transcription factor 1) Verhoeven AJM
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 73
Functional domains of the USF1 protein. The A1 domain is important for E-box dependent transactivation, the USR (USF-
specific region) and A2 domains are important for E-box and initiator element (Inr)-dependent transactivation (Roy et al., 1997). Post-translational modifications that affect USF1 function are indicated. The protein product of the splice variant lacks the first 59
amino acids, dimerizes with full-length USF1 protein, which results in its inactivation (Saito et al., 2003).
Protein
Description
USF1 belongs to the bHLH-Zip class of
transcription factors. The bHLH-ZIP domains are
important for DNA binding and dimerization. USF
homo- and heterodimers activate transcription of
target genes through binding either at distal E-box
elements or at pyrimidine-rich Inr elements in the
core promoter (Roy et al., 1997). Whole genome
ChIP-chip analysis in human hepatoma HepG2
cells showed that USF1 and USF2 bind
predominantly to CACGTGAC elements (Rada-
Iglesias et al., 2008). In addition, USF2 but not
USF1 binds to pyrimidine rich elements, suggesting
that transactivation through Inr elements is mainly
through USF2. Transactivation activity critically
depends on post-translational modification of
USF1. DNA binding to the E-box element is
increased by phosphorylation of USF1 by the cdk1,
p38 stress-activated kinase, protein kinase A and
protein kinase C pathway (Corre and Galibert,
2005), whereas phosphorylation through the
PI3Kinase pathway leads to loss of DNA binding
activity to the ApoAV promoter (Nowak et al.,
2005). Cellular stress stimuli such as DNA damage,
oxidative stress and heavy metal exposure, induce
p38-mediated phosphorylation at T153
and increased
USF1 transactivation activity. Upon increased
and/or prolonged stress exposure, USF1
phosphorylated at T153
becomes acetylated at K199
with concomitant loss of transactivation activity
(Corre et al., 2009). In fasting-refeeding cycles,
insulin increases the transactivation activity of
USF1 via DNA-PK mediated phosphorylation of
residue S262
and subsequent acetylation at K237
(Wong et al., 2009).
Expression
The USF1 gene is ubiquitously expressed (Sirito et
al., 1994).
Localisation
The USF1 protein is located in the nucleus.
Function
USF1 has been shown to play an important role in
transcriptional regulation of a huge number of
seemingly unrelated genes (Corre and Galibert,
2005; Rada-Iglesias et al., 2008), consistent with
the abundant distribution of E-box like elements in
the genome. Whole-genome ChIP analysis in
HepG2 cells identified 2518 USF1 binding sites in
chromatin context, of which 41 % were located
within 1 kb of a transcription start site (Rade-
Iglesias et al., 2008). USF1 binding signals strongly
correlate with target gene expression levels,
suggesting that USF1 plays an important role in
transcription activation. USF1 physically interacts
with histone modifying enzymes, transcription
preinitiation complex factors, coactivator and
corepressor proteins (Corre and Galibert, 2005;
Huang et al., 2007; Corre et al., 2009; Wong et al.,
2009). In addition, USF1 interacts with other
transcription factors to achieve cooperative
transcriptional activation of individual genes (Corre
and Galibert, 2005). USF1 also plays a crucial role
in chromatin barrier insulator function, in which
euchromatin regions are protected from
heterochromatin-induced gene silencing (Huang et
al., 2007). USFs recruit histone modifying enzymes
to the insulator element, which modify the adjacent
nucleosomes thereby maintaining chromatin in an
open state and preventing heterochromatin spread.
Similarly, USFs main function at enhancer
elements may be to render the adjacent region
accessible for binding of other, bona fide
transcription factors, by the recruitment of histone
modifying enzymes (Huang et al., 2007).
Tumor suppression: Several lines of evidence
support the hypothesis that USF1 may act as a
tumor suppressor. First, USF1 is involved in the
transcriptional activation of several tumor
suppressor genes (e.g. p53, APC, BRCA2, PTEN,
SSeCKS) (Corre and Galibert, 2005; Pezzolesi et
al., 2007; Bu and Gelman, 2007), and represses
expression of human telomerase reverse
transcriptase TERT (McMurray and McCance,
2003; Chang et al., 2005). Second, USF1 is
involved in cell cycle control (Cogswell et al.,
1995) and overexpression of USF1 slows G2/M
transition in thyrocytes and thyroid carcinoma cells
(Jung et al., 2007). Third, USF1 overexpression
leads to a strong reduction in cell proliferation in
Ha-Ras/c-Myc transformed fibroblasts (Luo and
Sawadogo, 1996). Fourth, USF1 transactivation
activity is completely lost in three out of six
transformed breast cell lines (Ismail et al., 1999).
Fifth, USF1 antagonizes some activities of the
oncoprotein c-Myc, possibly by competing for the
same DNA binding sites (Luo and Sawadogo, 1996;
McMurray and McCance, 2003). Definitive proof
USF1 (upstream transcription factor 1) Verhoeven AJM
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 74
that USF1 is a tumor suppressor protein, e.g.
showing that USF1 knockdown increases cell
proliferation and tumor formation, however, is still
missing. This proof may be hard to gain, as USF2
may compensate for USF1 loss, and USF2 appears
to have a broader antiproliferative function than
USF1 (Luo and Sadawogo, 1996; Sirito et al., 1998;
Vallet et al., 1998).
Homology
The USF1 gene is widely conserved with orthologs
identified in Ciona intestinalis and Drosophila
melanogaster.
Mutations
Note
Of the 121 SNPs in the USF1 gene collected in the
dbSNP database, only the rs4126997 T>C
polymorphism causes a non-synchronous mutation
(V15
A missense), but data on allele frequency or
functional effects are not available. The two SNPs
that are shown to be functional, rs2073658 A>G in
intron 7 (heterozygosity 0.296) and rs3737787 C>T
in the 3'-UTR (heterozygosity 0.309), are in almost
complete linkage disequilibrium. The minor allele
is accompanied by normal USF1 expression in
human muscle and fat tissue but loss of insulin-
induced upregulation of USF1 mRNA and known
USF1 target genes (Naukkarinen et al., 2005;
Naukkarinen et al., 2009), as well as reduced
insulin-mediated anti-lipolytic activity (Kantartzis
et al., 2007).
Implicated in
Carcinogenesis
Note
Given the suggestive evidence for a role of USF1 in
tumor suppression, one may anticipate that
carcinogenesis will evolve from loss of USF1
transactivation activity, either as a result of
mutations in the USF1 gene or of posttranslational
modification of USF1 protein. This has not been
reported yet. Alternatively, tumor suppressor genes
may lose responsivity to USF1 by mutations in the
DNA binding element or by changes in local DNA
methylation. This is exemplified by the observation
of a classic Cowden syndrome patient with early
onset breast cancer and reduced PTEN activity,
which appears to be due to a specific germline
mutation of an E-box element in the PTEN gene
and loss of USF1 binding (Pezzolesi et al., 2007).
Familial combined hyperlipidemia (FCHL)
Disease
FCHL is the most common genetic form of
hyperlipidemia and is associated with increased risk
of premature cardiovascular disease. Affected
persons characteristically show elevation of both
cholesterol and triglycerides in the blood, which is
due to increased VLDL and LDL levels. This is
often accompanied by elevated apoB100 and low
HDL levels, and a preponderance of small dense
LDL particles (Naukkarinen et al., 2006). FCHL is
genetically heterogeneous. One of the loci that is
linked to FCHL is 1q21-q23. Pajukanta et al. (2004)
showed that the dyslipidemia observed in FCHL is
linked to the USF1 gene. The disease is associated
with a common haplotype of non-coding SNPs
within the USF1 gene. Carriers of the risk allele
show lack of insulin-induced increase of USF1
expression in skeletal muscle and fat tissue
(Naukkarinen et al., 2009). As USF1 is involved in
regulation of numerous genes of glucose and lipid
metabolism (Corre and Galibert, 2005), non-
responsive USF1 expression may lead to increased
production and reduced metabolism of plasma
lipids and lipoproteins.
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Roy AL, Du H, Gregor PD, Novina CD, Martinez E, Roeder RG. Cloning of an inr- and E-box-binding protein, TFII-I, that interacts physically and functionally with USF1. EMBO J. 1997 Dec 1;16(23):7091-104
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Huang S, Li X, Yusufzai TM, Qiu Y, Felsenfeld G. USF1 recruits histone modification complexes and is critical for maintenance of a chromatin barrier. Mol Cell Biol. 2007 Nov;27(22):7991-8002
Jung HS, Kim KS, Chung YJ, Chung HK, Min YK, Lee MS, Lee MK, Kim KW, Chung JH. USF inhibits cell proliferation through delay in G2/M phase in FRTL-5 cells. Endocr J. 2007 Apr;54(2):275-85
Kantartzis K, Fritsche A, Machicao F, Stumvoll M, Machann J, Schick F, Häring HU, Stefan N. Upstream transcription factor 1 gene polymorphisms are associated with high antilipolytic insulin sensitivity and show gene-gene interactions. J Mol Med. 2007 Jan;85(1):55-61
Pezzolesi MG, Zbuk KM, Waite KA, Eng C. Comparative genomic and functional analyses reveal a novel cis-acting PTEN regulatory element as a highly conserved functional E-box motif deleted in Cowden syndrome. Hum Mol Genet. 2007 May 1;16(9):1058-71
Rada-Iglesias A, Ameur A, Kapranov P, Enroth S, Komorowski J, Gingeras TR, Wadelius C. Whole-genome maps of USF1 and USF2 binding and histone H3 acetylation reveal new aspects of promoter structure and candidate genes for common human disorders. Genome Res. 2008 Mar;18(3):380-92
Corre S, Primot A, Baron Y, Le Seyec J, Goding C, Galibert MD. Target gene specificity of USF-1 is directed via p38-mediated phosphorylation-dependent acetylation. J Biol Chem. 2009 Jul 10;284(28):18851-62
Naukkarinen J, Nilsson E, Koistinen HA, Söderlund S, Lyssenko V, Vaag A, Poulsen P, Groop L, Taskinen MR, Peltonen L. Functional variant disrupts insulin induction of USF1: mechanism for USF1-associated dyslipidemias. Circ Cardiovasc Genet. 2009 Oct;2(5):522-9
Wong RH, Chang I, Hudak CS, Hyun S, Kwan HY, Sul HS. A role of DNA-PK for the metabolic gene regulation in response to insulin. Cell. 2009 Mar 20;136(6):1056-72
This article should be referenced as such:
Verhoeven AJM. USF1 (upstream transcription factor 1). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):72-75.
Gene Section Mini Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 76
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
WNK2 (WNK lysine deficient protein kinase 2) Peter Jordan
Departamento de Genetica, Instituto Nacional de Saude Dr Ricardo Jorge, Avenida Padre Cruz, 1649-
016 Lisboa, Portugal (PJ)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Genes/WNK2ID41867ch9q22.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI WNK2ID41867ch9q22.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity Other names: KAIA302, KIAA1760, NY-CO-43,
P/OKcl.13, PRKWNK2, SDCCAG43
HGNC (Hugo): WNK2
Location: 9q22.31
Local order: The WNK2 gene is covered by BAC
clones RP11-370F5, RP11-480F4 and RP11-165J3
and is flanked by the NINJ1 (telomeric) and
C9orf129 (centromeric) genes.
DNA/RNA
Description
The human WNK2 gene is composed of 31 exons
spanning 136 Kbp on chromosome 9q22.31. The
promoter region contains a 700 bp CpG island
between 1103 bp and 396 bp upstream of the ATG
translation start codon. A second CpG island spans
exon 1 from 135 bp upstream of the ATG
translation start codon until 638 bp downstream of
the ATG and close to the end of exon 1.
Transcription
Two major alternative transcripts exist depending
on the terminal exon chosen.
One variant uses exons 1-30, has a coding sequence
of 6894 bp and yields WNK2(1-2297) (related to
clone KIAA1760, Acc. Nb. AB051547). The other
variant skips exon 30 and includes exon 31, has a
coding sequence of 6765 bp and yields WNK2(1-
2254) (related to clone Kaia302; Acc. Nb.
AK000694). Both terminal exons 30 and 31 carry
their own 3'-untranslated regions and
polyadenylation signals. In addition, there is
evidence for alternative splicing in other exons and
in a tissue-specific manner.
Pseudogene
None known.
Protein
Description
Amino acids: 2297 or 2254. Molecular Weight:
243000 Daltons. The WNK2 protein encodes a
cytoplasmic serine-threonine kinase that lacks a
lysine in subdomain II required for ATP-binding in
most protein kinases and instead uses an alternative
lysine in subdomain I. WNK kinase form a separate
family branch, most closely related to kinases
MEKK, Raf and PAK.
Human WNK2 gene structure. The gene spans 136 Kbp, contains 31 exons and localizes to chromosome 9q22.31. Exons (vertical boxes) and separating introns are shown in proportion to their sizes; however, intron scale differs from exon scale.
WNK2 (WNK lysine deficient protein kinase 2) Jordan P
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 77
Diagram of the WNK2 protein in scale. The sequence contains a catalytic domain near the N-terminus and a coiled coil
domain near the C-terminus. Except for three short homology regions shared with the three other human WNK kinases, no other functional domains are known. The two splicing variants WNK2(1-2297) and WNK2(1-2254) differ in the C-terminal protein
sequence.
Expression
WNK2 is preferentially expressed in heart, skeletal
muscle and brain but also in small intestine, colon
and liver. Loss of expression was reported in a large
percentage of human gliomas (Hong et al., 2007)
and grade II and III meningiomas (Jun et al., 2009)
due to extensive methylation in the CpG island at
the 5' end of the WNK2 gene. In contrast, promoter
methylation was rare in other tumor types. This
finding makes WNK2 a candidate tumor suppressor
gene in brain tumors.
Localisation
The subcellular localization of GFP-tagged WNK2
in HeLa cells was predominantly cytoplasmic. Part
of the endogenous WNK2 pool in HT29 colorectal
cells localized to the plasma membrane and
overexpression of a WNK2(1922-2156) that
contains the coiled-coil domain was targeted to the
plasma membrane.
Function
Human WNK2 modulates the activation level of
ERK1 and ERK2. Experimental depletion of
WNK2 or overexpression of a kinase-dead
WNK2K207M mutant led to increased phospho-
ERK1/2 levels when a basal ERK stimulation was
present but not, for example, in serum-free culture
conditions (Moniz et al., 2007). This increase in
ERK1/2 activation promoted cell cycle progression
through G1/S and sensitized cells to respond to
lower concentrations of EGF. From these data one
might predict that loss of WNK2 expression will
promote cell cycle progression in tumor cells.
Interestingly, WNK2 expression is silenced in a
significant percentage of human gliomas (Hong et
al., 2007) suggesting that this pathway may be used
in some tumor types to promote cell proliferation.
The molecular mechanism through which a
reduction in WNK2 expression can increase
ERK1/2 activation involves phosphorylation of
MEK1 at serine 298, a modification that increases
MEK1 affinity towards ERK1/2. Apparently,
WNK2 affects PAK1 activation via Rac1 and
PAK1 is the kinase responsible for MEK1 S298
phosphorylation (Moniz et al., 2008).
Homology
The catalytic domain of WNK2 is 90% identical to
WNK1, 91% identical to WNK3 and 81% identical
to WNK4. The remaining sequence of WNK2 has
little homology to other WNK members except for
three small WNK homology regions (Holden et al.,
2004; Moniz et al., 2007). These include an acidic
motif (residues 586-597) to which hereditary
mutations in WNK4 cluster (Wilson et al., 2001),
residues 1186-1261 without any recognizable motif,
and residues 1918-1988 including a coiled-coil
domain.
Mutations
Note
At present it is unclear whether the observed
somatic mutations have a functional impact on the
WNK2 protein or confer any selective advantage to
tumors cells.
WNK2 (WNK lysine deficient protein kinase 2) Jordan P
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 78
Tissue Histology/Type cDNA Protein Mutation Ref
Colorectal adenocarcinoma c.1964delC p.P655fs*2 Frameshift
deletion Greenman et al., 2007
Brain glioblastoma c.3799G>A p.A1267T Missense Parsons et al., 2008
Stomach adenocarcinoma c.4269delC p.S1424fs*5 Frameshift
deletion Greenman et al., 2007
Lung neuroendocrine
carcinoma c.5009G>A p.G1670E Missense
Greenman et al., 2007; Davies
et al., 2005
Lung adenocarcinoma c.6089G>T p.S2030I Missense Greenman et al., 2007; Davies
et al., 2005
Ovary serous carcinoma c.1528G>T p.V510L Missense Greenman et al., 2007
Ovary mucinous carcinoma c.6798delC p.T2267fs*31 Frameshift
deletion Greenman et al., 2007
Germinal
No germinal mutations described.
Somatic
Somatic mutations in WNK2 have been found in
the course of large scale tumor genome sequencing
efforts (see Table above).
Heterozygous somatic mutations in the WNK2 gene
identified by large-scale tumor sequencing.
Implicated in
Brain tumors
Note
Promoter methylation leads to loss of expression.
Disease
Glioma and meningioma.
Prognosis
Unknown.
Colon cancer
Note
WNK2 clone was isolated as a serologically defined
colon cancer antigen 43; WNK2 is expressed in
colon.
To be noted Possible role in invasion due to the effect of WNK2
on Rho-GTPases. WNK2 controls (through a yet
unknown mechanism) the activation of RhoA,
which in turn determines the activation of Rac1 in a
reciprocal manner. Experimental depletion of
WNK2 leads to reduced RhoA and increased Rac1
activation.
References Scanlan MJ, Chen YT, Williamson B, Gure AO, Stockert E, Gordan JD, Türeci O, Sahin U, Pfreundschuh M, Old LJ. Characterization of human colon cancer antigens recognized by autologous antibodies. Int J Cancer. 1998 May 29;76(5):652-8
Ito M, Shichijo S, Tsuda N, Ochi M, Harashima N, Saito N, Itoh K. Molecular basis of T cell-mediated recognition of pancreatic cancer cells. Cancer Res. 2001 Mar 1;61(5):2038-46
Wilson FH, Disse-Nicodème S, Choate KA, Ishikawa K et al. Human hypertension caused by mutations in WNK kinases. Science. 2001 Aug 10;293(5532):1107-12
Holden S, Cox J, Raymond FL. Cloning, genomic organization, alternative splicing and expression analysis of the human gene WNK3 (PRKWNK3). Gene. 2004 Jun 23;335:109-19
Davies H, Hunter C, Smith R, Stephens P, et al. Somatic mutations of the protein kinase gene family in human lung cancer. Cancer Res. 2005 Sep 1;65(17):7591-5
Greenman C, Stephens P, Smith R, Dalgliesh GL, Hunter C, et al. Patterns of somatic mutation in human cancer genomes. Nature. 2007 Mar 8;446(7132):153-8
Hong C, Moorefield KS, Jun P, Aldape KD, Kharbanda S, Phillips HS, Costello JF. Epigenome scans and cancer genome sequencing converge on WNK2, a kinase-independent suppressor of cell growth. Proc Natl Acad Sci U S A. 2007 Jun 26;104(26):10974-9
Moniz S, Veríssimo F, Matos P, Brazão R, Silva E, Kotelevets L, Chastre E, Gespach C, Jordan P. Protein kinase WNK2 inhibits cell proliferation by negatively modulating the activation of MEK1/ERK1/2. Oncogene. 2007 Sep 6;26(41):6071-81
Moniz S, Matos P, Jordan P. WNK2 modulates MEK1 activity through the Rho GTPase pathway. Cell Signal. 2008 Oct;20(10):1762-8
Parsons DW, Jones S, Zhang X, Lin JC, Leary RJ, et al. An integrated genomic analysis of human glioblastoma multiforme. Science. 2008 Sep 26;321(5897):1807-12
Jun P, Hong C, Lal A, Wong JM, McDermott MW, Bollen AW, Plass C, Held WA, Smiraglia DJ, Costello JF. Epigenetic silencing of the kinase tumor suppressor WNK2 is tumor-type and tumor-grade specific. Neuro Oncol. 2009 Aug;11(4):414-22
Moniz S, Jordan P. Emerging roles for WNK kinases in cancer. Cell Mol Life Sci. 2010 Apr;67(8):1265-76
This article should be referenced as such:
Jordan P. WNK2 (WNK lysine deficient protein kinase 2). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):76-78.
Leukaemia Section Mini Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 79
Atlas of Genetics and Cytogenetics in Oncology and Haematology
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t(1;2)(p36;p21) Jean-Loup Huret
Genetics, Dept Medical Information, University of Poitiers, CHU Poitiers Hospital, F-86021 Poitiers,
France (JLH)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Anomalies/t0102p36p21ID1542.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI t0102p36p21ID1542.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Clinics and pathology
Disease
Myelodysplastic syndrome (MDS) in most cases,
acute lymphoblastic leukemia (ALL) in one case.
Phenotype/cell stem origin
At least 3 of the 6 available cases were treatment
related myelodysplastic syndromes (t-MDS)
(Roulston et al., 1998; Mauritzson et al., 2002;
Masuya et al., 2002), and 2 other cases were MDS
(Horiike et al., 1988; Storlazzi et al., 2008).
Clinics
A 38-year-old male patient presented with a
treatment related myelodysplastic syndrome (t-
MDS) evolving towards an acute myeloid leukemia
(t-AML). Previous treatment included
topoisomerase inhibitors for a Hodgkin disease 36
months before diagnosis of the t-MDS (Roulston et
al., 1998). A t-MDS was diagnosed in a 76-year-old
female patient previously treated with radiotherapy
for uterine cancer 29 years ago. She died 26 months
after diagnosis of the t-MDS (Mauritzson et al.,
2002). A 49-year-old female patient was diagnosed
with t-MDS (FAB refractory anemia (RA)); she had
been treated with etoposide 2 years previously for
M1-AML; the patient died 6.5 years after onset of
the t(1;2). Other chromosome anomalies appeared
during course of the disease, as well as an unrelated
clone (Masuya et al., 2002). A 67-year-old female
patient had a chronic myelomonocytic leukemia
(CMML) with a normal karyotype; she received
hydroxyurea. Three years later, a refractory anemia
with excess of blasts-2 (RAEB-2) and a t(1;2) was
diagnosed. The patient died one month later
(Storlazzi et al., 2008).
Refractory anemia with excess of blasts (RAEB)
was diagnosed in a 69-year-old male patient. The
patient was still alive 15 months after diagnosis
(Horiike et al., 1988). A T-cell acute lymphoblastic
leukemia (T-ALL) was found in a 1-year-old child
(Mathew et al., 2001).
Cytogenetics
Cytogenetics morphological
In two cases, the t(1;2) was the sole anomaly
(Horiike et al., 1988; Storlazzi et al., 2008). In
contrast, complex karyotype were present in the 4
other cases. inv(14)(q11q32) was present in the T-
ALL case (Mathew et al., 2001); del(5q) was found
in two cases (Roulston et al., 1998; Mauritzson et
al., 2002) and del(7q) in one case (Masuya et al.,
2002). Other remarkable anomalies were:
t(14;21)(q22;q22) with RUNX1 involvement
(Roulston et al., 1998), +8, +12, +13 appearing
during course of the disease (Masuya et al., 2002);
there was also, in the latter case, an unrelated clone
with t(11;12)(p15;q13).
Genes involved and proteins
Note
In only one case were the genes involved in the
translocation studied (Storlazzi et al., 2008).
PRDM16
Location
1p36
Protein
Transcription activator; PRDM16 forms a
t(1;2)(p36;p21) Huret JL
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 80
transcriptional complex with CEBPB. PRDM16
plays a downstream regulatory role in mediating
TGFB signaling (Bjork et al., 2010). PRDM16
induces brown fat determination and differentiation
(Kajimura et al., 2010).
FLJ42875
Location
1p36
DNA/RNA
2 transcript variants; non-coding RNA of unknown
function.
Result of the chromosomal anomaly
Fusion protein
Description
PRDM16 (both long and short isoforms) and
FLJ42875 are overexpressed. The sequence on
chromosome 2 upregulating these 2 genes is
unknown.
References Horiike S, Taniwaki M, Misawa S, Abe T. Chromosome abnormalities and karyotypic evolution in 83 patients with myelodysplastic syndrome and predictive value for prognosis. Cancer. 1988 Sep 15;62(6):1129-38
Roulston D, Espinosa R 3rd, Nucifora G, Larson RA, Le Beau MM, Rowley JD. CBFA2(AML1) translocations with novel partner chromosomes in myeloid leukemias: association with prior therapy. Blood. 1998 Oct 15;92(8):2879-85
Mathew S, Rao PH, Dalton J, Downing JR, Raimondi SC. Multicolor spectral karyotyping identifies novel translocations in childhood acute lymphoblastic leukemia. Leukemia. 2001 Mar;15(3):468-72
Masuya M, Katayama N, Inagaki K, Miwa H, Hoshino N, Miyashita H, Suzuki H, Araki H, Mitani H, Nishii K, Kageyama S, Minami N, Shiku H. Two independent clones in myelodysplastic syndrome following treatment of acute myeloid leukemia. Int J Hematol. 2002 Feb;75(2):182-6
Mauritzson N, Albin M, Rylander L, Billström R, Ahlgren T, Mikoczy Z, Björk J, Strömberg U, Nilsson PG, Mitelman F, Hagmar L, Johansson B. Pooled analysis of clinical and cytogenetic features in treatment-related and de novo adult acute myeloid leukemia and myelodysplastic syndromes based on a consecutive series of 761 patients analyzed 1976-1993 and on 5098 unselected cases reported in the literature 1974-2001. Leukemia. 2002 Dec;16(12):2366-78
Storlazzi CT, Albano F, Guastadisegni MC, Impera L, Mühlematter D, Meyer-Monard S, Wuillemin W, Rocchi M, Jotterand M. Upregulation of MEL1 and FLJ42875 genes by position effect resulting from a t(1;2)(p36;p21) occurring during evolution of chronic myelomonocytic leukemia. Blood Cells Mol Dis. 2008 May-Jun;40(3):452-5
Kajimura S, Seale P, Kubota K, Lunsford E, Frangioni JV, Gygi SP, Spiegelman BM. Initiation of myoblast to brown fat switch by a PRDM16-C/EBP-beta transcriptional complex. Nature. 2009 Aug 27;460(7259):1154-8
Bjork BC, Turbe-Doan A, Prysak M, Herron BJ, Beier DR. Prdm16 is required for normal palatogenesis in mice. Hum Mol Genet. 2010 Mar 1;19(5):774-89
This article should be referenced as such:
Huret JL. t(1;2)(p36;p21). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):79-80.
Leukaemia Section Short Communication
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 81
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
t(2;18)(q11;q21) Jean-Loup Huret
Genetics, Dept Medical Information, University of Poitiers, CHU Poitiers Hospital, F-86021 Poitiers,
France (JLH)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Anomalies/t0218q11q21ID2158.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI t0218q11q21ID2158.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Clinics and pathology
Disease
Non Hodgkin lymphoma.
Phenotype/cell stem origin
One case to date, a 65-year-old female patient with
a follicular lymphoma stage II-a (Impera et al.,
2008).
Evolution
Complete remission was obtained.
Cytogenetics
Additional anomalies
A complex karyotype was found, with +11, and
other anomalies.
Genes involved and proteins
AFF3
Location
2q11.2
Protein
AFF3 belongs to a family of putative transcription
factors also comprising AFF1 (AF4, FEL, MLLT2)
in 4q21, AFF2 (FMR2, FRAXE) in Xq28 and
AFF4 (AF5Q31) in 5q31. AFF3 has been found a
susceptibility gene in autoimmune diseases, namely
rheumatoid arthritis, psoriatic arthritis, and juvenile
idiopathic arthritis (Barton et al., 2009; Castelino
and Barton, 2010; Hinks et al., 2010). AFF3 is
deleted in Nievergelt syndrome, an autosomal
dominant mesomelic dysplasia (Steichen-Gersdorf
et al., 2008). AFF3 was also found expressed in
20% of mammary tumor cells but not in normal
acini in a study (To et al., 2005).
BCL2
Location
18q21.33
Protein
Antiapoptotic protein.
Result of the chromosomal anomaly
Hybrid gene
Description
Fusion of AFF3 exon 1 to BCL2 exon 2.
Fusion protein
Oncogenesis
Leads to the overexpression of BCL2.
References To MD, Faseruk SA, Gokgoz N, Pinnaduwage D, Done SJ, Andrulis IL. LAF-4 is aberrantly expressed in human breast cancer. Int J Cancer. 2005 Jul 1;115(4):568-74
Impera L, Albano F, Lo Cunsolo C, Funes S, Iuzzolino P, Laveder F, Panagopoulos I, Rocchi M, Storlazzi CT. A novel fusion 5'AFF3/3'BCL2 originated from a t(2;18)(q11.2;q21.33) translocation in follicular lymphoma. Oncogene. 2008 Oct 16;27(47):6187-90
Steichen-Gersdorf E, Gassner I, Superti-Furga A, Ullmann R, Stricker S, Klopocki E, Mundlos S. Triangular tibia with fibular aplasia associated with a microdeletion on 2q11.2 encompassing LAF4. Clin Genet. 2008 Dec;74(6):560-5
Barton A, Eyre S, Ke X, Hinks A, Bowes J, Flynn E, Martin P, Wilson AG, Morgan AW, Emery P, Steer S, Hocking LJ, Reid DM, Harrison P, Wordsworth P, Thomson W, Worthington J. Identification of AF4/FMR2 family, member
t(2;18)(q11;q21) Huret JL
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 82
3 (AFF3) as a novel rheumatoid arthritis susceptibility locus and confirmation of two further pan-autoimmune susceptibility genes. Hum Mol Genet. 2009 Jul 1;18(13):2518-22
Castelino M, Barton A. Genetic susceptibility factors for psoriatic arthritis. Curr Opin Rheumatol. 2010 Mar;22(2):152-6
Hinks A, Eyre S, Ke X, Barton A, Martin P, Flynn E, Packham J, Worthington J, Thomson W. Association of the AFF3 gene and IL2/IL21 gene region with juvenile idiopathic arthritis. Genes Immun. 2010 Mar;11(2):194-8
This article should be referenced as such:
Huret JL. t(2;18)(q11;q21). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):81-82.
Leukaemia Section Short Communication
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 83
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
t(2;21)(q11;q22) Jean-Loup Huret
Genetics, Dept Medical Information, University of Poitiers, CHU Poitiers Hospital, F-86021 Poitiers,
France (JLH)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Anomalies/t0221q11q22ID1551.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI t0221q11q22ID1551.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Clinics and pathology
Disease
T-cell acute lymphoblastic leukemia.
Phenotype/cell stem origin
One case to date, a 6-year-old boy (Chinen et al.,
2008).
Evolution
Complete remission was obtained. An allogenic
bone marrow transplantation was performed, and
the patient had remained in complete remission for
17 months at the time of the report.
Cytogenetics
Additional anomalies
A complex karyotype was found.
Genes involved and proteins
AFF3
Location
2q11.2
Protein
AFF3 belongs to a family of putative transcription
factors also comprising AFF1 (AF4, FEL, MLLT2)
in 4q21, AFF2 (FMR2, FRAXE) in Xq28 and
AFF4 (AF5Q31) in 5q31. AFF3 has been found a
susceptibility gene in autoimmune diseases, namely
rheumatoid arthritis, psoriatic arthritis, and juvenile
idiopathic arthritis (Barton et al., 2009; Castelino
and Barton, 2010; Hinks et al., 2010). AFF3 is
deleted in Nievergelt syndrome, an autosomal
dominant mesomelic dysplasia (Steichen-Gersdorf
et al., 2008). AFF3 was also found expressed in
20% of mammary tumor cells but not in normal
acini in a study (To et al., 2005).
RUNX1
Location
21q22.3
Protein
Transcription factor (activator) for various
hematopoietic-specific genes.
Result of the chromosomal anomaly
Hybrid gene
Description
Fusion of RUNX1 exon 7 to AFF3 exon 8.
References To MD, Faseruk SA, Gokgoz N, Pinnaduwage D, Done SJ, Andrulis IL. LAF-4 is aberrantly expressed in human breast cancer. Int J Cancer. 2005 Jul 1;115(4):568-74
Chinen Y, Taki T, Nishida K, Shimizu D, Okuda T, Yoshida N, Kobayashi C, Koike K, Tsuchida M, Hayashi Y, Taniwaki M. Identification of the novel AML1 fusion partner gene, LAF4, a fusion partner of MLL, in childhood T-cell acute lymphoblastic leukemia with t(2;21)(q11;q22) by bubble PCR method for cDNA. Oncogene. 2008 Apr 3;27(15):2249-56
Steichen-Gersdorf E, Gassner I, Superti-Furga A, Ullmann R, Stricker S, Klopocki E, Mundlos S. Triangular tibia with fibular aplasia associated with a microdeletion on 2q11.2 encompassing LAF4. Clin Genet. 2008 Dec;74(6):560-5
Barton A, Eyre S, Ke X, Hinks A, Bowes J, Flynn E, Martin P, Wilson AG, Morgan AW, Emery P, Steer S, Hocking LJ, Reid DM, Harrison P, Wordsworth P, Thomson W, Worthington J. Identification of AF4/FMR2 family, member
t(2;21)(q11;q22) Huret JL
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 84
3 (AFF3) as a novel rheumatoid arthritis susceptibility locus and confirmation of two further pan-autoimmune susceptibility genes. Hum Mol Genet. 2009 Jul 1;18(13):2518-22
Castelino M, Barton A. Genetic susceptibility factors for psoriatic arthritis. Curr Opin Rheumatol. 2010 Mar;22(2):152-6
Hinks A, Eyre S, Ke X, Barton A, Martin P, Flynn E, Packham J, Worthington J, Thomson W. Association of the AFF3 gene and IL2/IL21 gene region with juvenile idiopathic arthritis. Genes Immun. 2010 Mar;11(2):194-8
This article should be referenced as such:
Huret JL. t(2;21)(q11;q22). Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):83-84.
Solid Tumour Section Mini Review
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 85
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
Liver: Nested stromal epithelial tumor Y Albert Yeh
North Shore University Hospital, Long Island Jewish Medical Center, Hofstra University School of
Medicine, New York, USA (YAY)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Tumors/NestStromEpithLiverID6243.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI NestStromEpithLiverID6243.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity
Alias
Ossifying malignant mixed epithelial and stromal
tumor; Ossifying stromal-epithelial tumor;
Desmoplastic nested spindle cell tumor; Calcifying
nested stromal-epithelial tumors of the liver
Note
Nested stromal epithelial tumor (NSET) of the liver
is an extremely rare non-hepatocytic tumor of the
liver and is characterized by nests of spindle and
epithelioid cells with occasional calcification and
ossification.
Synonyms include ossifying malignant mixed
epithelial and stromal tumor, ossifying stromal-
epithelial tumor, desmoplastic nested spindle cell
tumor, and calcifying nested stromal-epithelial
tumors of the liver.
Clinics and pathology
Epidemiology
Twenty four cases of NSET of the liver have been
reported. NSET occurs in patients with age range
from 2 to 33 years old. The tumor affects mainly
children, and females are more frequently affected
than males.
Gross and histopathological characteristics of NSET. Grossly, the tumor has a yellow-tan and bulging lobulated appearance. A satellite tumor nodule is present in the periphery (A). Microscopically, nested tumor cells have oval nuclei, stippled chromatin,
and inconspicuous nucleoli. The interface between epithelioid and spindle cells is shown (arrows). Courtesy of Sergey V Brodsky.
Liver: Nested stromal epithelial tumor Yeh YA
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 86
Clinics
Patients are presented with palpable abdominal
tumors. Cushing syndrome has been reported in a
few patients.
Pathology
Macroscopically, NSETs predominantly occur in
the right lobe of the liver and are unencapsulated,
well circumscribed tumors range in size from 2.8 to
30 cm in greatest dimension. Satellite tumor
nodules also have been reported. Tumors have a
yellow-tan and lobulated appearance.
Microscopically, NSETs are characterized by
organoid arrangement of cellular nests composed of
spindle and epithelioid cells embedded in a
desmoplastic or fibrocytic/myofibroblastic stroma,
within which proliferation of bile ductules is noted.
Areas of myxoid and cystic degeneration or
necrosis are sometimes encountered within or
adjacent to the cellular nests. Focal psammomatous
calcification or osteoid formation is present in some
tumors. Lymphovascular invasion is occasionally
seen.
The spindle cells within the cellular nests are
arranged in short fascicles with a somewhat
whorled pattern.
The nested cells are characterized by plump nuclei,
stippled chromatin, and inconspicuous nucleoli.
Scattered mitotic figures with abnormal forms are
identified.
Immunohistochemically, the tumor cells are stained
positive for cytokeratin AE1/AE3, keratin CK19
(focal), EMA, CD117 (c-kit), CD56, CD99, ACTH,
chromogranin, synaptophysin, neuron-specific
enolase, and S100 (focally weak in epithelioid
cells). Vimentin stain is positive in the nested
spindled cell and stroma. Muscle specific actin and
smooth muscle actin immunostains highlight
stromal myofibroblastic cells. Alpha-fetoprotein
and p53 are negative.
Cytogenetics
Chromosomal analysis of one tumor reveals an
abnormal karyotype of 60-63,XXX,-1,-4,-5,-
others,+2mar.
Prognosis
Most patients are doing well with no tumor
recurrence in 6 months to 14 years. Tumor
recurrence has been observed in two of twenty four
patients. One case with aggressive clinical behavior
and extrahepatic lymph node metastasis has been
reported.
Immunohistochemical stains of NSET. Tumor cells are stained positive for immunostains including ACTH (A) (Courtesy of
Milton J Finegold), b-catenin (B), cyclin D1 (C), Ki-67 (30% of nuclear staining) (D), p21ras (E), topoisomerase II (F).
Liver: Nested stromal epithelial tumor Yeh YA
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 87
References Ishak KG, Goodman ZD, Stocker JT.. Miscellaneous malignant tumors. In: Rosai J, Sobin LH, eds. Tumors of the liver and intrahepatic bile ducts. Armed forces Institute of Pathology, Washington DC, 2001:271-2.
Heywood G, Burgart LJ, Nagorney DM. Ossifying malignant mixed epithelial and stromal tumor of the liver: a case report of a previously undescribed tumor. Cancer. 2002 Feb 15;94(4):1018-22
Heerema-McKenney A, Leuschner I, Smith N, Sennesh J, Finegold MJ. Nested stromal epithelial tumor of the liver: six cases of a distinctive pediatric neoplasm with frequent calcifications and association with cushing syndrome. Am J Surg Pathol. 2005 Jan;29(1):10-20
Hill DA, Swanson PE, Anderson K, Covinsky MH, Finn LS, Ruchelli ED, Nascimento AG, Langer JC, Minkes RK, McAlister W, Dehner LP. Desmoplastic nested spindle cell tumor of liver: report of four cases of a proposed new entity. Am J Surg Pathol. 2005 Jan;29(1):1-9
Brodsky SV, Sandoval C, Sharma N, Yusuf Y, Facciuto ME, Humphrey M, Yeh YA, Braun A, Melamed M, Finegold MJ. Recurrent nested stromal epithelial tumor of the liver
with extrahepatic metastasis: case report and review of literature. Pediatr Dev Pathol. 2008 Nov-Dec;11(6):469-73
Makhlouf HR, Abdul-Al HM, Wang G, Goodman ZD. Calcifying nested stromal-epithelial tumors of the liver: a clinicopathologic, immunohistochemical, and molecular genetic study of 9 cases with a long-term follow-up. Am J Surg Pathol. 2009 Jul;33(7):976-83
Meir K, Maly A, Doviner V, Gross E, Weintraub M, Rabin L, Pappo O. Nested (ossifying) stromal epithelial tumor of the liver: case report. Pediatr Dev Pathol. 2009 May-Jun;12(3):233-6
Grazi GL, Vetrone G, d'Errico A, Caprara G, Ercolani G, Cescon M, Ravaioli M, Del Gaudio M, Vivarelli M, Zanello M, Pinna AD. Nested stromal-epithelial tumor (NSET) of the liver: a case report of an extremely rare tumor. Pathol Res Pract. 2010 Apr 15;206(4):282-6
Oviedo Ramírez MI, Bas Bernal A, Ortiz Ruiz E, Bermejo J, De Alava E, Hernández T. Desmoplastic nested spindle cell tumor of the liver in an adult. Ann Diagn Pathol. 2010 Feb;14(1):44-9
This article should be referenced as such:
Yeh YA. Liver: Nested stromal epithelial tumor. Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):85-87.
Deep Insight Section
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 88
Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
Inhibitors of the heat shock protein 90: from cancer clinical trials to neurodegenerative diseases Jean François Peyrat, Samir Messaoudi, Jean Daniel Brion, Mouad Alami
Université Paris-Sud, CNRS, BioCIS-UMR 8076, Laboratoire de Chimie Thérapeutique, Faculté de
Pharmacie, 5 rue J.-B. Clément, Châtenay-Malabry, F-92296, France (JFP, SM, JDB, MA)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Deep/HSP90inCancerTreatmentID20086.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI HSP90inCancerTreatmentID20086.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
I) Introduction
The 90-kDa heat shock protein 90, Hsp90, belongs
to the family of molecular chaperone responsible
for the conformational maturation or reparation of
other proteins, referred to as "clients", into
biologically active structures (Pearl and Prodromou,
2006). Hsp90 exerts its essential ATP dependant
chaperone function to more than three hundred
client proteins involved in cell growth,
differentiation and survival (Workman, 2004;
Chiosis et al., 2004; Sreedhar et al., 2004b; Zhang
and Burrows, 2004; Neckers, 2002). Many of them,
more than forty, include overexpressed or mutant
oncogenic proteins ErbB2/HER2 (Miller et al.,
1994; An et al., 1997), Braf (Grbovic et al., 2006),
Akt/PKB (Sato et al., 2000), muted p53
(Blagosklonny et al., 1996), transcription factors:
hormone steroid receptors GR (Grad and Picard,
2007) ER and AR, angiogenic factors HIF-1α
(Picard, 2006; Kuduk et al., 2000; Kuduk et al.,
1999; Johnson and Toft, 1995), telomerase
(Forsythe et al., 2001; Akalin et al., 2001) which
are associated with the six hallmarks of cancer
(Figure 1).
Under non-stress conditions the quaternary
structure of Hsp90 is now well established to be a
dimeric complex, and its abundance is
approximately 1% of the total protein contents.
Each monomer consists in three domains: the N-
terminal domain (NTD), a middle domain (MD)
implicated in client protein binding, and a C-
terminal dimerization domain (CTD) (Figure 2)
(Harris et al., 2004; Shiau et al., 2006).
Figure 1: Hsp90 protein partners and clients destabilized by Hsp90 inhibition (Jackson et al., 2004).
Inhibitors of the heat shock protein 90: from cancer clinical trials to neurodegenerative diseases
Peyrat JF, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 89
Figure 2: Structure of the full length yeast Hsp90, in complex with non hydrolysable ATP analogue (Hsp90 ATP) and structure of
the full length E. coli Hsp90 without nucleotide (Hsp90 apo) (Shiau et al., 2006).
In the human proteome, several isoforms of Hsp90
have been isolated, Hsp90α (inducible form) and
Hsp90β (constitutive form) localized in cytoplasm
(Sreedhar et al., 2004a), while Grp94 (glucose-
regulated protein) and TRAP-1 (HSP75/tumor
necrosis factor receptor associated protein 1) are
localized in the endoplasmic reticulum and in
mitochondria respectively (Csermely et al., 1998;
Maloney and Workman, 2002). To acquire its full
active molecular chaperone activity, Hsp90
operates with molecular co-chaperones and partner
proteins to form a series of multimeric protein
complexes (Figure 3) including Hsp70, peptidyl-
prolyl isomerases, immunophilins (FKBP51 and
FKBP52) and the cyclophilin CYP40. Others co-
chaperones such as p23, recently identified as a
prostaglandine E2-Syntase, plays an important role
in the activity of a number of transcription factors
of the steroid/thyroid receptor family (Chan et al.,
2008; Grad et al., 2006).
It is now well established, that Hsp90 needs to bind
ATP in a pocket located in the N-terminal domain
to exert its function. Thus, the Hsp90 protein
function may be inhibited by molecules competing
with ATP binding (such as geladanamycin: GA,
Figure 3), thereby freezing the chaperone cycle,
which in turn decreases the affinity of Hsp90 for
client proteins and leads to 26S proteasome-
mediated oncogenic client protein degradation
(Sepp-Lorenzino et al., 1995). N-terminal domain
Hsp90 inhibitors block cancer cell proliferation in
vitro and cancer growth in vivo (Sharp and
Workman, 2006).
To date, the full crystal structure of Hsp90 in
complex with a non hydrolysable ATP analogue
(Ali et al., 2006), and of the full length E.Coli
Hsp90 without nucleotide (apo-Hsp90) (Shiau et
al., 2006), have yet been reported (Figure 2).
Furthermore, an interesting recent study
investigated Hsp90 conformational changes in
solution, shows a long range effects between Hsp90
domains, as the binding of co-chaperones (or
inhibitors) at NTD induce conformational changes
in the MD and CTD (Phillips et al., 2007). The C-
terminal domain has been implicated biochemically
as the site of a possible second, cryptic ATP-
binding site on Hsp90. Its contribution to the
overall regulation of chaperone function is not
clear, but the antibiotic novobiocin (Nvb) (c.f.
structure in Figure 15) has been reported to bind
this site and alter the conformation of the chaperone
(Yun et al., 2004).
Since pharmacological inhibition of Hsp90 by
several families of small molecules leading to the
degradation of oncogenic proteins, Hsp90 has
become a target of interest against cancer and
allowed the development of numerous small
inhibitors (Biamonte et al., 2010).
II) Hsp90 health and cancer (Powers and
Workman, 2007)
Hsp90 has probably been most widely
acknowledged as a therapeutic target for the
treatment of cancer (Mitsiades et al., 2007).
Although there is no evidence of Hsp90 mutations
in malignancy, there is increasing support for the
view that this molecular chaperone plays an
important role in the development, maintenance and
progression of cancers.
One of the principal debates concerning the
inhibition of the highly abundant Hsp90 is the
selectivity of inhibitors for the chaperone protein in
malignant cells (Kamal et al., 2003). Some works
suggest that Hsp90 inhibitors could provide an
exploitable therapeutic index (Banerji, 2005).
Firstly, it has been reported that inhibitors were
significantly more sensitive to Hsp90 in cancer
cells (Neckers and Neckers, 2005; Powers and
Workman, 2006; Chiosis, 2006; Whitesell et al.,
1994; Neckers, 2006). In support to this surprising
observation, Kamal et al. showed that the activity
state of the Hsp90 chaperone machine was different
in tumor cells.
Inhibitors of the heat shock protein 90: from cancer clinical trials to neurodegenerative diseases
Peyrat JF, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 90
Figure 3: Hsp90 cycle: GA: geldanamycin analogue; 40=Hsp40; 70=Hsp70; IP: immunophillin; HIP: Hsp70-interacting protein; HOP=Hsp70/Hsp90 organizing protein (Biamonte et al., 2010).
Indeed, the Hsp90 is entirely bound in an active
complex with co-chaperones, whereas most Hsp90
in normal tissues resides in a free, uncomplexed
state (Workman, 2004; Kamal et al., 2003).
Furthermore, Hsp90 is constitutively expressed at
higher levels (2-10 fold) in tumor cells compared
with their normal counterparts. This higher Hsp90
activity is probably due to the
overexpression/amplification of mutated Hsp90
clients, and this is in correlation with the higher
level of cochaperones of Hsp90 observed in
cancerous cells.
Finally, the selective sensitivity of transformed
cells for Hsp90 inhibitors may be partly due to the
selective accumulation of these drugs in cancer
cells since the in vivo observation of Hsp90
inhibitors in murine model system showed higher
concentration in tumor tissue (Chiosis and Neckers,
2006).
Consequently, the Hsp90 has emerged as an
exciting target for the development of cancer
chemotherapeutics. However, despite the numerous
molecules which have prompted a phase I clinical
trial, it remains to be verified if Hsp90 inhibitors
will provide adequate treatment in clinic.
III) Hsp90 inhibitors (Messaoudi, 2008)
The Hsp90 protein function may be inhibited with
molecules that bind the ATP pocket, or its
chaperone activity may be disturbed by small
molecules binders interfering with domains in the
C-terminus or median region. Although Hsp90
function provides an attractive target for the
treatment of cancer, the feasibility and efficacy of
the inhibitors approach has just begun to be
explored in clinic.
Direct inhibitors of Hsp90 have been divided into
two groups:
A) N-terminal domain binders
1) Ansamycin macrolactames
1.a) Quinone derivatives
Geldanamycin (Figure 4), was isolated from the
broth of Streptomyces hygroscopicus in 1970s (De
Boer et al., 1970). Further studies have shown that
GA revert the phenotype of v-src oncogene
transformed cells. However, this ability was not due
to a direct action of the Src kinase activity, but to an
inhibition of Hsp90. Subsequent
immunoprecipitation and X-ray cristallographic
studies have shown that GA competes with ATP
and binds to the N-terminal domain site of Hsp90,
leading the Hsp90 multichaperone complexes to the
ubiquitin-mediated proteasome degradation (Roe et
al., 1999; Stebbins et al., 1997). Since this
observation, GA was used to identify additional
Hsp90 substrates and to understand the role of
Hsp90 in promoting malignant transformation.
Although GA provided very promising antitumor
effects, it showed several pharmacologic limitations
as poor solubility, limited in vivo stability and high
hepatotoxicity in some of the human tumor models
(Neckers, 2006; Supko et al., 1995). Thus, the 17-
position of GA has been an attractive focal point for
the synthesis of GA analogues. Structure-activity
relationship (SAR) studies have shown that
structurally and sterically diverse 17-substituents
Inhibitors of the heat shock protein 90: from cancer clinical trials to neurodegenerative diseases
Peyrat JF, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 91
can be introduced without destroying antitumor
activity. Then, further derivatives of GA, with
similar biological behaviour but a better toxicity
profile, were synthesized (Schulte and Neckers,
1998). Therefore, new C-17 substituted derivatives
17-AAG (17-allyl-17-desmethoxygeldanamycin,
also designed KOS953, CNF 1010, tanespimycin,
Figure 4) and 17-DMAG (17-(2-
dimethylaminoethylamino)-17-
desmethoxygeldanamycin, KOS1022,
alvespimycin, Figure 4) (Snader et al., 2002; Solit
et al., 2007) were brought to the fore by displaying
a significant enhancement of the
chemical/metabolic stability.
17-AAG can be used in single agent or in
combination with other cancer therapeutics
(KOS953/bortezomib (Anderson, 2007; Richardson
et al., 2007), KOS953/trastuzumab (Modi et al.,
2007), 17-AAG/Paclitaxel (Sain et al., 2006), 17-
AAG/cisplatin (McCollum et al., 2008)).
To enhance the pharmacokinetics and dynamics of
17-AAG, Kosan Biosciences Incorporated has
developed a DMSO-free formulation (KOS953)
contained cremophor, which is actually in Phase I
clinical testing.
Although 17-AAG and its numerous formulations
have shown some encouraging clinical responses,
they present important drawbacks (e.g.; liver
toxicity and cumbersome formulation) that may
limit their clinical applications whereas 17-DMAG
exhibits a better water solubility and oral
bioavailability (Ronnen et al., 2006). However,
although clinical trials in myeloid leukemia seemed
to be promising, the 17-DMAG was discontinued in
2008 (ClinicalTrials.gov).
1.b) Hydroquinone derivatives
In a different approach, Infinity Pharmaceuticals
has developed IPI504 (retaspimycin or 17-AAG
hydroquinone, Figure 4) (Adams et al., 2005; Sydor
et al., 2006), a new GA analogue, in which the
quinone moiety was replaced by a dihydroquinone
one. Indeed, the preclinical data suggested that the
hepatotoxicity of 17-AAG was attributable to the
ansamycin benzoquinone moiety, prone to
nucleophilic attack. Furthermore, it was recently
reported that the hydroquinone form binds Hsp90
with more efficiency than the corresponding
quinone form (Maroney et al., 2006). In biological
conditions, the hydroquinone form can interconvert
with GA, depending on redox equilibrium existing
in cell. It has been recently proposed, that NQ01
(NAD(P)H: quinone oxidoreductase) can produce
the active hydroquinone from the quinone form of
IPI504 (Chiosis, 2006). However, Infinity
Pharmaceuticals showed that if the overexpression
of NQ01 increased the level of hydroquinone and
cell sensitivity to IPI504, it has no significant effect
on its growth inhibitory activity. These results
suggest that NQ01 is not a determinant of IPI504
activity in vivo (Douglas et al., 2009).
1.c) Clinical trials
In 2007, results of the phase I clinical trial of
tanespimicyn (KOS953) with bortezomib in
patients with relapsed refractory multiple myeloma
were reported (Solit and Chiosis, 2008; Taldone et
al., 2008). Dose escalations in the trial ranged from
100 to 340 mg/m2 for tanespimycin, and from 0.7 to
1.3 mg/m2 for bortezomib. Results showed that two
patients, on the 41 enrolled, exhibited stable disease
after two cycles, and 18 of them demonstrated a
response to combination (Richardson et al., 2007).
Moreover, the tanespimicyn was co-administrated
with trastuzumab on 25 patients treated with up to
450 mg/m2 of drug on a weekly schedule. This
combination induced a regression of 21, 22 and
25% in three patients, which had failed trastuzumab
therapy, with HER2-amplified breast cancer (Modi
et al., 2007).
Figure 4: GA, 17-AAG, 17-DMAG and IPI504.
Inhibitors of the heat shock protein 90: from cancer clinical trials to neurodegenerative diseases
Peyrat JF, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 92
In 2008, Infinity Pharmaceuticals reported the
results of a dose escalation Phase I/II clinical trial
of retaspimycin hydrochloride in patients with
metastatic and/or unresectable gastrointestinal
stromal tumors (GIST) on a twice weekly schedule
(400 mg/m2). 4 of the 18 patients enrolled, achieved
a partial response and 11/18 achieved stable
disease. These results had initiated the phase III
clinical trial of the study in 2008. However, Infinity
Pharmaceuticals reported on April 2009, the
decision to end its phase III study (RING trial) of
IPI504 hydrochloride in patients with refractory
gastrointestinal stromal tumors (Infinity Press
Release). The trial was based on 46 patients whose
tumors persist despite treatment with Gleevec
(imatinib) and Sutent (sunitinib). Resulting data
showed a higher than anticipated mortality rate. In
this heavily pretreated population, IPI504 was not
tolerated (400 mg/m2 or placebo in 21 days cycles
as a 30 min intravenous infusion twice weekly for 2
weeks followed by a 1 week rest) and the study was
terminated early.
Nevertheless, the IPI504 is still evaluating in phase
II trials in patients with non-small cell lung cancer,
and in combination with herceptin (trastuzumab) in
patients with HER-2 positive metastatic breast
cancer.
In the same month, KOSAN, acquired by BMS in
2008, reported that the phase III clinical trial
concerning the KOS953 or tanespimycin, in
combination with Bortezomib in patients with
multiple myeloma in first relapse has been
suspended. This was probably a precaution as the
metabolization of tanespimycin leads to IPI504.
Conforma Therapeutics/Biogen idec developed a
hydroquinone form of the 17-AAG (CNF1010),
trapped as HCl salt, which was in clinical phase I
against chronic lymphocytic leukemia. However, to
date, this program is terminated. Moreover, the
CNF1010 had started a phase III trial against GIST
in 2008. This study was also suspended due to the
anticipated mortality rate of patients enrolled
(ClinicalTrials.gov). Parallel efforts to improve the
solubility and bioavailability of 17-AAG have led
the NCI and Kosan to develop 17-DMAG
(KOS1022) as a second generation alternative
which has entered Phase I clinical testing (Santi et
al., 2007). Promising results were obtained in
patients with chemotherapy refractory acute
myelogenous leukemia, as 3 of 17 patients had a
complete response to therapy (Lancet et al., 2006).
However, researches were given up in 2008, as the
17-DMAG presents an unusable toxicity profile.
1.d) Other analogues
Diverse derivatives of 17-AAG bearing non-redox-
active phenol group designed by Kosan Biosciences
were reported (Tian et al., 2007). Amongst them,
KOSN1559 was claimed as the most potent Hsp90
inhibitor (e.g.; SKBr3 Cell Line IC50=860 nM,
Kd=16 nM) (Figure 5). To date, no clinical trial had
been reported with this compound.
Figure 5: Structure of KOSN 1559.
2) Purines
2.a) Purines analogues
Limitations in the clinical use of 17-AAG and 17-
DMAG have prompted the discovery of novel
Hsp90 ATPase inhibitors with improved "drug-
like" structural characteristics and better
pharmacological profiles. To this end, structure-
based design and high-throughput screening
approaches performed at the Memorial Sloane
Kettering Institute, have been taken to identify new
chemotypes that inhibit Hsp90 ATPase activity. A
significant breakthrough in the preparation of
synthetic Hsp90 inhibitor was the PU3 (Figure 6).
On the basis of X-ray analysis and molecular
modelling, Chiosis's group, showed that PU3 was
designed to place the purine moiety into the same
spatial orientation as adenine ring of ATP in the
nucleotide pocket of Hsp90 (Chiosis et al., 2001).
PU3 presented molecular signature of Hsp90
inhibition, including the degradation of HER2, even
if its affinity for Hsp90 is moderate.
Chiosis's group and Conforma therapeutics/Biogen
Idec optimized this class of compounds leading to
new analogues bearing a thioether bridge to connect
the purine nucleus to substituted phenyl rings.
Among them, the PUH58 (Figure 7) (Llauger et al.,
2005), an 8-arylsulfanyl analogue of PU3, has been
identified as the most potent and selective purine.
Further efforts in optimization of this lead
compound led to the development of PU24F-Cl,
(Figure 7) which presents a higher affinity (30
times more than PU3) for the N-terminus of the
Hsp90, and low micromolar activity in a cell
proliferation assay (Chiosis et al., 2002; Vilenchik
et al., 2004). In an in vivo experiment in MCF-7
tumor bearing mice, PU24FCl led to 70% inhibition
when administered at a dose of 200 mg/kg every
second day for 30 days (Vilenchik et al., 2004).
Inhibitors of the heat shock protein 90: from cancer clinical trials to neurodegenerative diseases
Peyrat JF, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 93
Figure 6: Co-crystal structures of ADP (left, in green) and PU3 (right, in magenta) with Hsp90.
Figure 7: Structures of PU24FCl and analogues.
Additional investigations concerning the
pharmacophore of this family were done. It was
demonstrated that the presence of an amino group
on the C2 position of the purine nucleus respects
the global size of the molecule in regard to the
parent one (PU3). In addition, it offers multiple
possibilities for hydrogen bonding, and thus
allowed the connection of the benzyl group on the
N-9 rather than the C-8 of the purine. Thus,
Conforma-Biogen idec have identifed the BIIB021
(originally named CNF 2024, Figure 7) (Kasibhatla
et al., 2007) which displayed a binding affinity of
1.7 nM (4.6 nM for 17-AAG) and induces
degradation of HER2 with an IC50 of 38 nM in
MCF-7 cells (Lundgren et al., 2009). BIIB021
compound was entered in clinical trials in 2005.
2.b) Clinical trial
Currently, BIIB021 is the only purine analogue
evaluated in phase I/II clinical trials in combination
with trastuzumab (herceptin) against breast cancer,
with an aromatase inhibitor (exemestane) in
metastatic HER2-advanced breast cancer, or alone
in subjects with gastrointestinal stromal tumors.
Results from the phase I trials showed that BIIB021
was well tolerated (800 mg twice weekly) and
induced a significant inhibition of the HER2 (Elfiky
et al., 2008).
3) Pyrazole and isoxazole derivatives
3.a) Pyrazole analogues
In 2004, CCT018159 (Figure 8), the first Hsp90
inhibitor in the pyrazole series, was identified by
Workman et al. from a library of 60000
compounds, using a high throughput screening
(HTS), in the Cancer Research UK Centre for
Cancer Therapeutics (Rowlands, 2004). This
compound inhibits the N-terminal ATPase activity
of yeast and human Hsp90 with an IC50 of 7.1 and
3.2 µM, respectively (Cheung et al., 2005; Sharp et
Inhibitors of the heat shock protein 90: from cancer clinical trials to neurodegenerative diseases
Peyrat JF, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 94
al., 2007a). Further HTS studies undertaken by
Genomics institute of the Novartis Research
Foundation (GNF) and based on time-resolved
fluorescence resonance energy transfer (TR-FRET)
had allowed identifying two leads, G3129 and
G3130 (Figure 8), amongst the one million
molecules screened (Kreusch et al., 2005).
However, both compounds exhibited relatively poor
binding affinity to N-terminal domain of the Hsp90
(Kd=680 and 280 nM respectively). In SkBr3 breast
cancer cells, G3130 caused the degradation of
HER2 (IC50=30 µM) while G3129 was ineffective.
In addition, the co-crystal structures of G3129 and
G3130, bound to the N-terminus of human Hsp90α,
were reported (Kreusch et al., 2005). From this
study, it was showed that the resorcinol ring bound
Hsp90 in a similar way than that of radicicol, a
resorcylic lactone that inhibits Hsp90.
Figure 8: Structure of analogues G3129 and G3130.
Figure 9: Co-crystal structure of G3130 bound to the N-terminus of human Hsp90α.
Figure 10: VER49009 and VER50589.
Inhibitors of the heat shock protein 90: from cancer clinical trials to neurodegenerative diseases
Peyrat JF, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 95
Furthermore, the 5-ethyl appendage projected into
the aromatic pocket that accommodates the benzyl
group of the purine analogues described before. The
pyrazole provides hydrogen bound acceptor, and
the imidazole (G3130) occupies the same pocket as
the quinone of GA (Figure 9).
Based on these data, medicinal chemistry efforts led
to the identification of the more potent analogue of
CCT018159, the VER49009 (IC50 ATPase
activity=0.14 µM, Figure 10), where the amide
group was a key in forming a new interaction with
the residue Gly97 of the protein (Dymock et al.,
2005).
Synta Pharmaceuticals Corp. had reported another
class of triazoles analogues as modulators of Hsp90
(Figure 11) (Ying et al., 2009). It has been shown
that STA-9090, an unspecified new resorcinol-
containing triazole compound (Lin et al., 2008),
inhibits the activity of Hsp90 protein from 10 to
100 µM and thereby leading to degradation of
Hsp90 client proteins such as HER2 gene product
(Ying et al., 2009). More recently, it had been
shown that the STA-1474, a highly soluble
phosphate prodrug of STA-9090, exhibits very
interesting biologic activity against osteosarcoma
cell lines (McCleese et al., 2009).
Figure 11: Pyrazoles reported by Synta Pharmaceuticals.
3.b) Isoxazole analogues
Further optimization of potency, pharmakinetic and
pharmacodynamic properties of VER49009, were
undertaken by Vernalis Ltd. (in collaboration with
Novartis) to offer a series of isoxazole resorcinol
inhibitors. One of these was the VER50589 (Figure
10) which exhibited a higher affinity (Kd=5 nM)
than VER49009 (Kd=78 nM) (Sharp et al., 2007b).
Thus, the pyrazole to isoxazole switch does not
affect the critical hydrogen bound of the pyrazole
resorcinol unit that anchors this class of inhibitors
to the Hsp90 NH2-terminal ATP site (Brough et al.,
2008). Moreover, VER50589 also showed
improved cellular uptake over VER49009.
Brough et al. from Vernalis, reported recently the
identification of new diarylisoxazole compound
VER52296 (Figure 12) (Brough et al., 2008; Eccles
et al., 2008). The areas of interest for SAR studies
were the 5' positon on the resorcinol ring, and the
para substitution of the phenyl group on the
isoxazole ring. It has been shown with the X-ray
cristal structure, that the replacement of the
chlorine, in regard to VER50589, by an isopropyl
group, results in an additional hydrophobic
interaction with Leu107 in the flexible lipophilic
pocket of the N-terminal site of Hsp90. Additional
hydrophobic interactions were also observed with
Thr109 and Gly135 from the morpholine moiety
present in VER52296/NVP-AUY922 (Figure 12).
This latter, subsequently developed by Novartis,
was found to be very potent in the Hsp90
Fluorescence Polarization binding assay (IC50=21
nM) and displays an average GI50 of 2-40 nM in
antiproliferation assays against different human
tumor cell lines (Brough et al., 2008). In addition,
as evaluated by cassette dosing to mice bearing
subcutaneous HCT116 human colon cancer,
VER52296/NVP-AUY922 was retained in HCT116
xenograft tumors when administered i.p., at
concentrations well above the GI50. Further in vivo
characterization in a human colon cancer xenograft
model, VER52296/NVP-AUY922, also inhibits
tumor growth by ~50% when dosed at 50 mg/kg i.p.
daily. Moreover VER52296/NVP-AUY922 induces
the degradation of HER2 with an IC50 of 7 nM. In
addition, VER52296/NVP-AUY922 was tested in
several xenografts (colon, glioblastoma, breast,
ovarian, prostate) and a therapeutic response was
observed in each case (Cheung et al., 2005; Jensen
et al., 2008; Eccles et al., 2008).
Figure 12: Structure of VER52296/NVP-AUY922.
3.c) Clinical trials
The VER52296/NVP-AUY922 is the sole isoxazole
analogue currently in phase I/II clinical trials as a
single agent or in combination with bortezomib or
dexamethasone, in patients with relapsed or
refractory multiple myeloma. AUY922 is also used
as a single agent, in advanced solid malignancies
and efficacy in HER2+ or ER
+ locally advanced or
metastatic breast cancer patients. In this latter phase
I/II trial, VER52296/NVP-AUY922 was
intravenously administrated once a week schedule.
The maximum dose reported is 54 mg/m2. At 40
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mg/m2, VER52296 induces an up regulation (4-19
fold) of Hsp70 and a 20% reduction in soluble
HER2 was achieved by 74% of patients.
Noteworthy, the STA9090 (Figure 11) is currently
enrolling patients in several phase I/II clinical trials
against solid tumor, myeloid leukemia, non-small
cell lung cancer and gastrointestinal stromal tumor.
4) Dihydroindazolone derivatives
4.a) SNX2112 and SNX5422
In 2007, Serenex Inc./Pfizer had started a phase I
clinical trial program for the SNX5422 (Figure 13).
Using a new screening platform, the compounds
retained on the ATP-affinity column were analyzed
by mass spectrometry leading to the identification
of a poorly soluble analogue, SNX2112, which will
become the glycine prodrug SNX5422. The
SNX2112 showed higher activity to bind Hsp90
(Ki=1 nM) than that of 17-DMAG and induced the
degradation of HER2 with an IC50 of 20 nM. Based
on the observation that breast cancer cell lines with
HER2 amplification are more sensitive to 17-AAG,
the SNX2112 was tested using a panel of breast,
lung, and ovarian cancer cell lines. In all cell lines
studied, SNX2112 inhibited cell proliferation with
IC50 ranging from 10 to 50 nmol/L. In contrast to
17-AAG, the sensitivity of cancer cell lines to
SNX2112 in vitro did not correlate with the level of
HER2 expression (Chandarlapaty et al., 2008;
Huang et al., 2006). This compound uniformly
targets both the pro-proliferation pathways driven
by HER2 and ERK as well as the anti-apoptotic Akt
pathway (Okawa et al., 2009). Indeed, it exhibits
potent in vivo antitumor activity that extends
significantly the effects observed with GA
analogues.
4.b) Clinical trial
SNX-5422 is currently in a phase I clinical trial in
treating patients with solid tumor or lymphoma that
has not responded to treatment. SNX5422 is equally
tested to treat solid tumor cancer and lymphomas,
and, in subjects with refractory hematological and
solid tumor malignancies. In 2008, results of a
phase I dose-escalation study of SNX5422 reported
that this compound was well tolerated at 21 mg/m2.
5) Other inhibitors
Another class of ATP Hsp90 inhibitors bearing a
resorcinol moiety is radicicol (Rd) (Figure 14), a
natural resorcylic lactone, isolated from the fungus
Monocillium nordinii and Monosporium bonorden.
Rd, also known as monorden, has been described to
reverse the Src-transformed morphology of
fibroblast (Whitesell et al., 1994). This effect was
first attributed to the inhibition of the oncogenic Src
(v-Src), and later proved to act as an inhibitor of
Hsp90 despite its difference in structure to GA.
Moreover, Rd was found to compete with GA for
binding to the NTD of the chaperone, suggesting
that Rd shares the geldanamycin binding site. This
compound is a potent and specific inhibitor of the
ATPase activity of Hsp90 with nanomolar affinity
(Kd=19 nM). This causes destabilization of Hsp90
client proteins (v-Src, Raf-1, ErbB2 and Ras), many
of which are essential for tumor cell growth.
Although, the in vitro antitumoral activity of Rd is
very promising however, its in vivo activity is very
weak probably because of its chemical instability in
serum and its rapid conversion into inactive
metabolites due to the electrophilic nature of the
dienone moiety.
Therefore, synthetic efforts have been directed to
identify radicicol derivatives with improved in vivo
activity (Proisy et al., 2006). To date, Kyowa Hakko
described novel oxime-derivatives of Rd, including
KF55823 and KF25706 (Soga et al., 2003) (Figure
14). Although these compounds exhibit potent
antitumor activities in preclinical models and do not
seem to cause hepatotoxicity, their clinical
evaluations of these compounds has not been
pursued.
Figure 13: Structure of SNX5422 and SNX2112.
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Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 97
Figure 14: Resorcylic inhibitors.
Figure 15: Structures of novobiocin, chlorobiocin and coumermycin A1.
B) C-terminal domain binders
In 1991, Csermely, Kahn and co-workers reported
the presence of a C-terminal ATP binding site on
Hsp90 which becomes accessible when the N-
terminal Bergerat pocket is occupied (Sõti et al.,
2002). A decade later, it has been shown that Nvb
interacts with an ATP-binding domain in the C-
terminus of Hsp90 (Marcu et al., 2000a).
Biochemical studies on the CTD of Hsp90 have
identified an allosteric regulation process with the
N-terminus site, where the occupancy of one site
blocks the interaction of the ligand with the other
site (Garnier et al., 2002). The structure of the full
length and middle and C-terminal construction of
Hsp90 with different nucleotide states (apo, ATP)
have shown that there is a hinge region between the
middle and C-terminal region of Hsp90. The
conformation of this region is dictated by the status
of the nucleotide at the N-terminal site. This
observation is in accordance with the allosteric
regulation of ATP binding. It suggests that the
putative secondary ATP site could be located at the
immediate proximity of the hinge.
IV) Coumarin inhibitors
Coumarin group antibiotics, such as novobiocin
(Nvb), coumermycin A1 (Kd=10 nM) and
clorobiocin (Figure 15), are potent inhibitors of the
bacterial ATP binding gyrase B, a type II DNA
topoisomerase (Gormley et al., 1996). Their affinity
for gyrase is considerably higher than that of
modern fluoroquinolones. These antibiotics have
been isolated from various Streptomyces species
(Lanoot et al., 2002) and all possess a 3-amino-4-
hydroxycoumarin moiety as a key structural feature.
Nvb is licensed as an antibiotic for clinical use
(Albamycin; Pharmacia-Upjohn) and for the
treatment of infections with multi-resistant gram-
positive bacteria such as Staphylococcus aureus and
S. epidermidis (Raad et al., 1995; Raad et al., 1998;
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Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 98
Rappa et al., 2000). It had been demonstrated that
the interaction of Nvb with Hsp90 induces
alteration in the affinity of the chaperone for GA
and Rd and causes in vitro and in vivo depletion of
key regulatory Hsp90-dependant kinases including
v-Src, Raf-1 and ErbB2 (e.g., ErbB2 in SkBr3
breast cancer cells ~700 µM). In addition, Nvb was
found to bind the C-terminal nucleotide binding
region of Hsp90, albeit with a lower affinity than
with gyrase B. Moreover, Nvb disrupts the
interaction of both the cochaperones p23 and
HSC70 with the Hsp90 complex.
In 2005, the first attempt to improve the inhibitory
activity of Nvb against Hsp90 was reported (Yu et
al., 2005; Blagg et al., 2006). These authors have
highlighted the crucial role of the noviose moiety at
the 7-position of the coumarin ring for the
biological activity. Compound A4 lacking the 4-
hydroxyl of the coumarin moiety and containing an
N-acetyl side chain in lieu of the benzamide was the
most active compound. This compound was
identified as Hsp90 inhibitor that induced
degradation of Hsp90-dependent client proteins at
70-fold lower concentration than Nvb. Recently, in
continuation of their structural modification studies,
the same authors reported that 3'-descarbamoyl-4-
deshydroxynovobiocin DHN2 (Figure 16) and
compound KU135 (Shelton et al., 2009) proved to
be a more effective and selective Hsp90 inhibitor
(degradation of ErbB2 and p53 between 0.1 and 1.0
µM) (Burlison et al., 2006).
Our group reported a novel series of 3-
aminocoumarin analogues (Le Bras et al., 2007a;
Le Bras et al., 2007b) lacking the noviose moiety as
a class of highly potent Hsp90 inhibitors. A
representative example of this new class of
inhibitors is 4TCNA (Figure 17) (Le Bras et al.,
2007b).
In these analogues, the introduction of a tosyl
substituent on C-4 position of coumarin nucleus
(4TCNA) contributed to a significant extent for
maximal activity despite weaker water solubility.
Moreover, this lead has a particular implication in
apoptotic process. Thus, 4TCNA promotes
apoptosis through activation of caspases 7 and 8 in
ER-positive MCF-7 human breast cancer cells,
whereas in Ishikawa endometrial adenocarcinoma
cells, it induced apoptosis that was associated with
caspase activation and cleavage of PARP.
Furthermore, characterization of its mode of action
revealed that 4TCNA induced-cleavage of the p23,
recently identified as a prostaglandine E2-Syntase,
which plays an important role in activity of a
number of transcription factors of steroids/thyroid
receptors family. These results demonstrate that this
new denoviose compound presents originality in
regard to Nvb osidic derivatives already known.
In another study based on a simplified 3-
aminocoumarin scaffold, we also demonstrated that
4-tosyl-7-deshydroxycyclonovobiocic acid
(4TDHCNA) (Figure 17) (Radanyi et al., 2008),
exhibit increased inhibitory activity against the
Hsp90 protein folding process (MCF7 IC50=50
µM).
This result shows that removal of C7/C8
substituents is not detrimental for Hsp90 inhibitory
activity and strongly enhances the capacity of
4DHTCNA to inhibit Hsp90. This compound was
identified to be the most potent representative of the
new family of simplified coumarins. Results from
this study suggest that 4TDHCNA and 4TCNA,
which exerted similar biological profile may be
considered interesting compounds for the
development of more potent novobiocin analogues.
More recently, results from our group allowed the
identification of a new family of novobiocin
analogues in which the coumarin unit has been
replaced by a 2-quinoleinone moiety (unpublished
results). The quinolone-scaffold represents a
platform for the creation of easily synthesizable
soluble molecules. Compound 4-tosyl-3[(chroman-
6-yl) carboxylamino]-2-quinolon (4TCCQ, Figure
17) (IC50=5-8 µM) is 100-fold more potent than the
parent natural compound (novobiocin) and 6-fold
more active than the synthetic analogue 4TCNA.
Additionally, 4TCCQ induces the degradation of
ERα and strongly induces the cell death in MCF-7
breast cancer cell line.
Overall, these data provides compelling evidence
for the continued development of novobiocin-based
C-terminal domain Hsp90 inhibitors as promising
alternative to N-terminal domain inhibitors.
Figure 16: Structures of A4, DHN2 and KU135.
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Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 99
Figure 17: Structures of 4TCNA, 4TDHCNA and 4TTCQ.
Figure 18: Tau-Hsp90 in AD.
V) HSP 90 in Alzheimer's disease
Hsp90, a molecular chaperone, has come into its
own as a tantalizing target for cancer therapies.
However, its important functions of stabilization,
rematuration, disaggregation of many client
proteins could be exploitable in others diseases.
Indeed, neurodegenerative diseases are
characterized by the accumulation of misfolded
proteins that results in plaque formation. These
proteins rely upon HSP's for their refolding and
viability. Recently, it was suggested that Hsp90
may play a crucial role in maintaining pathogenic
changes that lead to neurodegenerative diseases
(Luo et al., 2008). Furthermore, the inhibition of
Hsp90 by 17-AAG derivatives and geldanamycin,
induces the HSP induction via HSF-1 activation,
resulting in neuroprotective activities. In the
Alzheimer's disease, the most common tauopathy,
in addition to β-amyloid deposition, there is an
accumulation of abnormal species of
hyperphosphorylated protein tau which leads to the
formation of toxic neurofibrillary tangles (Luo et
al., 2007; Dickey et al., 2007). This
hyperphosphorylation is caused by abnormal
kinases (CdK4, GSK-3β) activities resulting in
dissociation of transformed tau from microtubules,
aggregation and formation of neurofibrillary tangles
which can block the synaptic transmission (Figure
18).
Thus, the decrease of hyperphosphorylated tau
levels through refolding or degradation may
provide a possible therapeutic strategy against AD.
In this purpose, Dickey and Luo have presented
evidence that the stability of p35, (neuronal
activator of CdK4) and P301L mutant (most
common mutation in Alzheimer disease) are
maintaining by Hsp90.
Dou et al. 2007 reported that Hsp90 associates with
GSK-3β, regulating its stability and function,
preventing its degradation by the proteasome and so
allowing the increase of tau hyperphosphorylation.
Thus, the use of Hsp90 inhibitors leads to a
destabilization of GSK-3β and to a decrease of
hyperphosphorylated tau protein.
Dickey et al. 2007 demonstrated that CHIP (a tau
ubiquitin ligase) is intimately linked to tau
degradation following Hsp90 inhibition and that
this process is specific for promoting degradation of
only aberrant phosphorylated tau due to the fact that
the Hsp90 complex, in AD brain, presents higher
affinity for inhibitors than in unaffected brain
tissue.
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Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 100
Recently, compound A4 (Figure 16) was found to
exhibit significant protection against the Aβ-
induced toxicity at low concentrations (Lu, 2009).
These results suggest that novobiocin analogues
may represent an effective class of novel
compounds for treatment of AD.
VI) Conclusion
Since the first discovery of natural analogues, GA
and RD, the search for inhibitors of Hsp90 has
generated considerable interest as evidenced by the
number of compounds in clinical evaluations (Table
1).
However, if the first clinical results were very
encouraging, it seems that currently the
development of Hsp90 inhibitors experiencing
some difficulties, especially due to their toxicity.
Stopping clinical trials of IPI504, which
represented the most advanced HSP90 inhibitors, is
the unfortunate illustration of that. Thus, many
efforts are still needed in the understanding of the
administration of these agents but also in the
synthesis of new molecules. Moreover, the
involvement of Hsp90 in other non-oncological
diseases such as Alzheimer's disease shows the
importance of acquiring new and more potent
inhibitors with suitable pharmacological and
pharmacokinetic profiles.
Table 1: Current clinical trials.
Condition: R: recruiting, ANR: active, not recruiting, C: completed, NYR: not yet recruiting, S: suspended, T: terminated.
Therapy: ADL: Advanced Dedifferentiated Liposarcoma, AGC: Advanced Gastric Cancer, AM: Advanced Malignancies, AML: Acute Myeloid Leukemia, AST: Advanced Solid Tumors, BC: Breast Cancer, CLL: B-Cell Chronic Lymphotic Leukemia, CLL:
Chronic Lymphocytic Leukemia, CML: Chronic Myelogenous Leukemia, ALL: Acute Lymphoid Leukemia, MM: Multiple Myeloma, MetM: Metastatic Myeloma, NSCLC: Non Small Cell Lung Cancer, PC: Prostate Cancer, GIST: Gastrointestinal Stroma Tumor,
RRMM: Relapsed or Refractory Multiple Myeloma, ST: Solid Tumors, UST: Unresectable Solid Tumors.
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This article should be referenced as such:
Peyrat JF, Messaoudi S, Brion JD, Alami M. Inhibitors of the heat shock protein 90: from cancer clinical trials to neurodegenerative diseases. Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):88-104.
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Atlas of Genetics and Cytogenetics in Oncology and Haematology
OPEN ACCESS JOURNAL AT INIST-CNRS
LDI-PCR in Cancer Translocation Mapping Björn Schneider, Hans G Drexler, Roderick AF MacLeod
DSMZ, German Collection of Microorganisms and Cell Cultures, Department of Human and Animal
Cell Cultures, Inhoffenstr. 7b, 38124 Braunschweig, Germany (BS, HGD, RAFM)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Deep/LDI-PCRinCancerID20087.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI LDI-PCRinCancerID20087.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Abstract Identification of genes in oncogenic chromosome translocations by Fluorescence In Situ Hybridization (FISH)
screening using genomic tilepath clones, is often laborious, notably if the region of interest is gene-dense. Other
molecular methods for partner identification also suffer limitations; for instance, genomic PCR screening
requires prior knowledge of both sets of breakpoints, while Rapid Amplification of cDNA Ends (RACE) is
limited to translocations causing mRNA fusion and delivers no breakpoint data. With Long Distance Inverse
(LDI)-PCR, however, it is possible to identify unknown translocation partners and to map breakpoints at the
base-pair level. Applying LDI-PCR merely requires approximate sequence information on one partner, rendering
it ideal for use in combination with FISH to extend and refine cytogenetic breakpoint data.
Introduction Recurrent chromosomal rearrangements
characterize many different types of cancer.
Specific cytogenetic translocations are key events,
widely considered to be diagnostically and
prognostically significant in leukemia and
lymphoma, and increasingly so in solid tumors
(Mitelman et al., 2007). Hitherto, most cancer
genes have been identified following analysis of
recurrent chromosome translocations (Futreal et al.,
2004). The pathological significance and usefulness
of such rearrangements depend on two key features:
a) whether rearrangements display distinct patterns
of recurrence within specific tumors, e.g.
t(8;14)(q24;q32) which is restricted to B-cell
neoplasia; and b) how clustered are the
chromosomal breakpoints therein. The significance
accorded to breakpoint data depends on their
ascertainment precision, from megabase- and
kilobase-, down to single base-pair levels, when
ascertained by classical cytogenetics, fluorescence
in situ hybridization (FISH), and sequence-based
methods, respectively.
Chromosome translocations fall into three broad
categories. The first causes the physical fusion of
the two mRNAs expressed by the participating
genes, thus creating novel fusion proteins translated
from exons emanating from both genes, e.g. BCR
(at [chromosome]-22-[band]-q11) with ABL1 (at
9q34) fused by t(9;22)(q34;q11) in chronic myeloid
leukemia (CML) and in some cases of acute
lymphoblastic leukemias (ALL) (Turhan, 2008a,b).
The second category also fuses mRNA from genes
at separate loci but, in this case, serving to
deregulate a developmentally silenced partner by
exchanging promoters with more active partners,
e.g. BCL6 (at 3q27) which is activated by
translocations with any one of many partners,
chiefly in diffuse large B-cell lymphoma (DLBCL)
(Knezevich, 2007).
The third class of chromosome translocation again
results in the activation of the normally silent
partner, this time by juxtaposition with another
constitutively active partner without mRNA fusion,
e.g. the neighboring homeobox genes, TLX3 (at
5q35.1) and NKX2-5 (at 5q35.2). According to the
proximity of the breakpoint involved, either (but
not both) genes may be activated in T-cell ALL by
the recurrent t(5;14)(q35;q32.2) by which these are
juxtaposed with regulatory regions from BCL11B
LDI-PCR in Cancer Translocation Mapping Schneider B, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 106
(at 14q32.2) to stimulate transcription (Bernard et
al., 2001; MacLeod et al., 2003; Nagel et al., 2007).
Some "promiscuous" genes engage with multiple
partners: notably MLL with 64 known partners
(Meyer et al., 2009b), BCL6 with 28 (Knezevich,
2007), RUNX1 with 39 (Huret and Senon, 2003),
NUP98 with 29 (Kearney, 2002), and the IgH-locus
with 40 (Lefranc, 2003).
Although, promiscuity may reflect the dependence
of tumors on the inappropriate oncogene expression
without overly caring how deregulation is
accomplished, the role of the partner genes has
come under renewed scrutiny. Choice of partner
gene may not only reveal in which types of
precancerous cells primary oncogenic
rearrangements occur, but also by looking for
conserved DNA or protein motifs, yield clues to the
mechanisms underlying their formation or their
functional contribution to neoplasia. In addition, the
roles of biologically important genes, e.g. BCL11B,
a key regulator of both differentiation and survival
during thymocyte development, are often first
rendered visible by their participation in cancer
rearrangements (MacLeod et al., 2003).
Both the identities of each partner gene and their
precise breakpoints at the DNA base-pair level,
may be useful not only to characterize potential
fusion genes/products but also to ascertain whether
additional non protein-coding genomic entities,
such as chromosomal fragile sites (Schneider et al.,
2008), microRNA loci, putative genes, unspliced
"expressed sequence tags", or regulatory non-
coding regions may be involved.
Clues to the biological mechanisms generating
chromosome rearrangements are given by
breakpoint sequences, including T- and B-cell
receptor gene (VDJ) rearrangement, Alu-mediated
recombination, non-homologous end joining, etc.
VDJ genes are flanked by recombination signal
sequences composed of heptamers followed, in
turn, by a spacer containing either 12 or 23
unconserved nucleotides and a conserved nonamer.
Spacers of 12 nucleotides undergo physiological
recombination with those containing 23 in order to
obey the so-called "12/23 rule". The presence of
these or related sequences has been reported in
connection with cancer translocations to reveal how
physiologic processes may be abused to cause
genomic rearrangements (Gu et al., 1992).
Genomic fusion sequences may also be used to aid
cell line authentication - an omnipresent problem
confronting cell culturists, given that an
unexpectedly (and unacceptably) high percentage of
new cell lines has been misidentified, or cross-
contaminated by older cell lines (MacLeod et al.,
1999). While mRNA fusion sequences are
constrained by splicing, their genomic equivalents
allow sufficient variation to provide "fingerprints"
unique to individual cell lines to serve as potential
identifiers. For analyzing patient tumors,
knowledge of the exact fusion sequence allows
design of patient-specific quantitative (q)PCR used
for monitoring minimal residual disease with high
sensitivity (Burmeister et al., 2006), to follow up
therapeutic responses thus enabling early detection
of relapse.
Figure 1: Amplifying Genomic Fusions of Unknown Sequence. The schema summarizes how the genomic DNA is first restricted, then re-ligated to the circular template, and how the resulting amplicon should appear. Note unknown region (red) flanked by
known sequences (black). R: restriction site, BP: breakpoint; arrows: forward (FW) and reverse (REV) primers.
LDI-PCR in Cancer Translocation Mapping Schneider B, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 107
Cancer gene promiscuity enables oncogenic
chromosome rearrangements, "smoking guns" of
cancer genes, to be distinguished from random
changes. FISH is initially used to confirm
rearrangement of a contextually appropriate
oncogene residing at the locus in question. Hence, a
breakpoint at 9q34 might throw suspicion onto
NOTCH1 in a T-cell-, ABL1 in myeloid- neoplasia,
and NUP214 in either entity. While such an
approach is less helpful among solid tumors where
oncogene rearrangements are less informative, at
this locus TSC1 might be deemed a candidate in
tuberous sclerosis cells. Even when the index
breakpoint is precisely known, determination of its
partner by FISH requires time-consuming and
laborious procedures for those not afforded
immediate blanket tilepath-clone coverage with
which to quarter the region of interest.
PCR screening with hit-lists of known and potential
partner genes is quite as laborious as FISH, and is
liable to miss unknown translocation partners, or
those with breakpoints lying outside their respective
cluster regions. When there are grounds to suspect
transcriptional fusion (as among partners of genes
prone to this type of gene rearrangement, e.g.
ABL1, ETV6, NUP98, etc.) a mRNA-based
method, rapid amplification of cDNA ends
(RACE), may be used to detect novel fusion
partners (Frohman et al., 1988). A drawback of
RACE is its inability to supply genomic breakpoint
data, and the risk of overlooking some splice
variants. Hence, the technique of choice for
identifying unknown partner genes and their
breakpoints should not require prior knowledge of
the partner gene, yet provide breakpoint data at the
DNA base pair level.
Long Distance Inverse (LDI)-PCR satisfies these
needs. LDI-PCR was developed from the earlier
inverse-PCR (Ochman et al., 1988) to allow the
amplification of large DNA fragments comprised of
known and unknown sequences (Willis et al., 1997)
using re-ligated circular restriction fragments as
templates. Primers are set in opposition within the
known sequence. The unknown sequence is flanked
on both sides by known sequences following re-
ligation in the resultant amplicon (Fig. 1). When a
restriction fragment length polymorphism (RFLP)
distinguishing the wild type and derivative alleles is
generated by the genomic alteration, the two
resulting amplicons should be separable by gel
electrophoresis, enabling their respective sequences
to be compared (Fig. 2). Sequencing with one of the
PCR-primers directed towards the restriction site
allows immediate identification of the partner gene.
Sequencing in the other direction allows precise
mapping of the breakpoint.
Figure 2: How to Interpret LDI-PCR Gels. Left figure shows the wild type configuration where twin circular templates identical in size would yield a single band by agarose gel electrophoresis. Translocation bearing cells (right figure) yield both wild type and
derivative templates, differing in size and detectible as two bands on the gel. The derivative band is indicated by an arrow. Known regions are outlined in black, unknown in red. R: Restriction site, REV: reverse primer, FW: forward primer.
LDI-PCR in Cancer Translocation Mapping Schneider B, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 108
While easier to perform for genes with well
defined, short breakpoint cluster regions, LDI-PCR
may be applied to any gene or region involved in a
translocation and has, therefore, been applied to a
wide variety of translocations involving both
frequently rearranged promiscuous oncogenes, but
also as single events. Table 1 gives an overview of
genes analyzed by LDI-PCR according to literature
databases and the respective references.
Limitations are set by the performance of the DNA
polymerase since lengthier fragments may resist
amplification, and by the placement of the RFLP, as
fragments similar in size cannot be readily
distinguished by gel electrophoresis. If primary
patient tumor material is analyzed, it should be
noted that the samples used for analysis not only
contain tumor material, but also normal bystander
cells devoid of tumor rearrangement. Detection
attempted at lower tumor infiltration rates risk false
negative results.
In contrast to other PCR methods suitable for
detection of unknown fusion sequences, such as
panhandle PCR (Megonigal et al., 2000) or
analogous techniques requiring adaptor ligations
(reviewed in Tonooka and Fujishima, 2009), LDI-
PCR is independent of any additional adaptors or
anchors which have to be ligated to the restricted
fragments, reducing the number of steps required,
while remaining sufficiently flexible to allow a
wide choice of restriction enzymes.
In the future, translocation analysis by next
generation sequencing should overcome these
limitations and suitable algorithms have been
developed to recognize novel derivative breakpoint-
flanking sequences and thereby identify novel
cancer translocations and other synonymous
rearrangements, including a subset of fusogenic
microdeletions (Campbell et al., 2008).
Methology In principle, LDI-PCR utilizes digested and re-
ligated circular templates, which are of different
sizes, due to RFLP caused by genomic
rearrangements. This size difference renders the
amplicons separable by gel electrophoresis (Fig. 2).
For a successful analysis, the LDI-PCR has to be
designed carefully. The sequence covering the
genomic region of interest should be selected from
a genome browser (ENSEMBL, UCSC, NCBI) and
then pasted into the query box of a restriction map
generator (BioEdit, multiple online tools: SMS,
RestrictionMapper). Restriction enzymes should be
chosen to yield fragments in a size range of 2-5 kb.
If using a double-digest strategy with enzymes
producing sticky ends, these ends must be
compatible. Ensure that both enzymes perform well
in the same buffer and at the same temperature.
Primer pairs have to be designed in such a way that
one primer is directed towards the restriction site,
the other one in the opposite direction (see Figures).
The sequence lying between the primer tails is not
subject to amplification, so the gap should not be
excessive, ideally 30-50 bp. A breakpoint lying
therein cannot be detected unless another primer
pair, e.g. at the other end of the restriction fragment
is used. For longer fragments (greater than 5 kb,
say) use of a primer set consisting of one forward
and multiple reverse primers (or vice versa) can be
helpful. The oligonucleotides should be ~30 bp
with a Tm ~65°C and a GC-content of 40-60%.
For LDI-PCR template preparation high quality
genomic DNA should be used, meaning high purity
(260/280 1.8-2.0 and 260/230 > 2) and high
integrity without degradation. One microgram of
DNA is then digested with 30-50 U of each
restriction enzyme in the presence of the
appropriate digestion buffer in a total volume of
100 μl for 3-4 h at the temperature suitable for the
chosen enzymes (mostly 37°C), followed by heat
inactivation (where applicable) and purification,
preferably with a column based purification kit.
Phenol / chloroform purification followed by
precipitation may also be performed, but residual
phenol can disturb downstream processes. To form
the circular templates, the restriction fragments are
then religated with 5 U T4 ligase overnight at 4-8°C
in a total volume of 80 μL, terminated by heat
inactivation. These conditions favor the desired
self-ligation.
The PCR is performed best using a PCR kit suitable
for amplification of long templates and using 5 μL
(62.5 ng) of the digested and re-ligated DNA. The
PCR products are analyzed by gel electrophoresis.
Discrepant bands not corresponding to the
calculated amplicon size may represent amplicons
of translocated fragments. These are excised from
the gel, purified and subjected to sequence analysis
and, unless artefacts, may reveal the translocation
partner and the exact breakpoint of the
rearrangement subject to analysis.
LDI-PCR in Cancer Translocation Mapping Schneider B, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 109
Gene No. of Partners References describing LDI-PCR analysis
ALK 14 (Allouche, 2010) Ma et al., 2000
ANTXR1 1 (Oberthuer et al., 2005) Oberthuer et al., 2005
API2 1 (Mathijs and Marynen, 2001) Dierlamm et al., 1999
BCL6 28 (Knezevich, 2007)
Akasaka H et al., 2000; Akasaka T et al., 2000; Kurata et
al., 2002; Akasaka et al., 2003; Chen et al., 2003;
Montesinos-Rongen et al., 2003; Chen et al., 2006;
Schneider et al., 2008
BRD4 1 (Collin, 2007) Haruki et al., 2005
CALL 1 (Frints et al., 2003) Frints et al., 2003
E2A 5 (Huret, 1997) Wiemels et al., 2002a
ETV6 28 (Knezevich, 2005) Wiemels et al., 1999a; Wiemels et al., 1999b; Wiemels
and Greaves, 1999; Wiemels et al., 2008
IGH 40 (Lefranc, 2003)
Willis et al., 1997; Willis et al., 1998; Nardini et al., 2000;
Satterwhite et al., 2001; Sonoki et al., 2001; Bichi et al.,
2002; Sanchez-Izquierdo et al., 2003; Sonoki et al., 2004;
Akasaka et al., 2007; Lenz et al., 2007; Souabni et al.,
2007; d'Amore et al., 2008; Ishizaki et al., 2008; Russell et
al., 2008; Vieira et al., 2008; Vinatzer et al., 2008; Nagel
et al., 2009; Russell et al., 2009; Yin et al., 2009; Hu et al.,
2010
let-7a-2, miR-100 1 (Bousquet, 2008) Bousquet et al., 2008
MLH1 1 (Meyer et al., 2009b) Meyer et al., 2009a
MLL 64 (Meyer et al., 2009b)
Blanco et al., 2001; Meyer et al., 2005; Teuffel et al.,
2005; Attarbaschi et al., 2006; Burmeister et al., 2006;
Matsuda et al., 2006; Meyer et al., 2006a; Meyer et al.,
2006b; Strehl et al., 2006; Burmeister et al., 2008;
Balgobind et al., 2009; Bueno et al., 2009; Burmeister et
al., 2009; Matsuda et al., 2009; Meyer and Marschalek,
2009; Cóser et al., 2010; De Braekeleer et al., 2010; Lee et
al., 2010
NOTCH1 2 (Suzuki et al., 2009) Suzuki et al., 2009
PAX5 6 (Strehl, 2005) An et al., 2008; An et al., 2009
PDGFRA 6 (Dessen, 2009) Cools et al., 2003
PDGFRB 20 (Vizmanos, 2005) Walz et al., 2007; Walz et al., 2009
RUNX1 / AML1 39 (Huret and Senon, 2003) Xiao et al., 2001; Wiemels et al., 2002b
Table 1: Genes involved in translocations analyzed by LDI-PCR, numbers of yet known translocation partner genes and
references wherein the analysis of the particular gene is described.
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Schneider B, Drexler HG, MacLeod RAF. LDI-PCR in Cancer Translocation Mapping. Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):105-113.
Case Report Section Paper co-edited with the European LeukemiaNet
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 114
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Unbalanced rearrangement der(9;18)(p10;q10) in a patient with polycythemia vera Xinjie Xu, Xueyan Chen, Elizabeth A Rauch, Eric B Johnson, Kate J Thompson, Jennifer
JS Laffin, Gordana Raca, Daniel F Kurtycz
University of Wisconsin-Madison, School of Medicine and Public Health, Department of Pediatrics,
University of Wisconsin Cytogenetic Services, Wisconsin State Laboratory of Hygiene, Madison, WI,
USA (XX, JJSL); University of Wisconsin-Madison, Department of Pathology and Laboratory
Medicine, Madison, WI, USA (XC); University of Wisconsin Cytogenetic Services, Wisconsin State
Laboratory of Hygiene, Madison, WI, USA (EAR, EBJ, KJT); University of Wisconsin-Madison,
School of Medicine and Public Health, Department of Pathology and Laboratory Medicine, University
of Wisconsin Cytogenetic Services, Wisconsin State Laboratory of Hygiene, Madison, WI, USA
(GR); University of Wisconsin-Madison, School of Medicine and Public Health, Department of
Pathology and Laboratory Medicine, Wisconsin State Laboratory of Hygiene, Madison, WI, USA
(DFK)
Published in Atlas Database: April 2010
Online updated version : http://AtlasGeneticsOncology.org/Reports/der0918XuID100044.html Printable original version : http://documents.irevues.inist.fr/bitstream/DOI der0918XuID100044.txt This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Clinics
Age and sex
69 years old female patient.
Previous history
No preleukemia ; no previous malignancy ; no
inborn condition of note.
Organomegaly
no hepatomegaly , splenomegaly , no enlarged
lymph nodes , no central nervous system
involvement (there was no apparent central nervous
system involvement at diagnosis).
Blood WBC : 15.2X 10
9/l
HB : 11.7g/dl
Platelets : 894X 109/l
Blasts : 0% peripheral
Bone marrow : 2% blasts
Cyto-Pathology Classification
Cytology
NA
Immunophenotype
NA
Rearranged Ig Tcr
NA
Diagnosis
Polycythemia vera
Survival
Date of diagnosis: 03-2005
Treatment: Phlebotomy
Complete remission : NA
Treatment related death : no
Relapse : NA
Status: Alive. Last follow up: 03-2010
Survival: 62 months
Karyotype
Sample: Bone marrow biopsy Sep 17th 2009.
Culture time: analysis was performed on overnight
colcemid and 24-hour cultures.
Banding: 400 band level.
Unbalanced rearrangement der(9;18)(p10;q10) in a patient with polycythemia vera
Xu X, et al.
Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1) 115
Results:
46,XX,+9,der(9;18)(p10;q10)[11]/46,XX[9]
Karyotype of a metaphase from the follow up specimen
from September 2009 (four years after the initial diagnosis), 24-hour culture.
FISH confirmation for an extra copy of 9p.
Comments
We describe a new case of der(9;18)(p10;q10)
detected in a patient with polycythemia vera (PV).
This rare rearrangement has been reported in five
cases of PV and one case of therapy associated
acute myeloid leukemia (t-AML) after essential
thrombocythemia (ET). Two of the five cases of PV
showed progression from PV to post-polycythemic
fibrosis, suggesting an association between this
cytogenetic abnormality and disease progression.
The patient presented in this report was diagnosed
with PV in 2005. Fluorescence In Situ
Hybridization (FISH) testing for the BCR/ABL
translocation was performed at diagnosis and was
negative. Subsequent molecular analysis detected
the presence of the JAK2 V617F mutation. The
patient had a bone marrow biopsy in September of
2009, due to worsening anemia which was at that
time attributed to excessive phlebotomy.
Cytogenetic analysis showed the presence of the
der(9;18)(p10;q10) in eleven out of twenty
analyzed cells. At the later follow-up visit in
February 2010, progression to the spent phase of
PV was suspected, based on the worsening of the
patient's clinical presentation. However, the next
bone marrow biopsy from March 2010 only
revealed mildly increased reticulin fibrosis. In
summary, although the patient currently does not
have pathohistological signs of progression,
clinically she is exhibiting worsening of the disease.
Gain of function of the JAK2 gene at 9p24 has a
crucial role in the pathogenesis of
myeloproliferative neoplasms. It has been proposed
that the der(9;18)(p10;q10) contributes to the
pathogenesis of PV through the gain of 9p, leading
to an extra copy of the JAK2 gene. For the patient
presented in this report the clinical significance of
the der(9;18)(p10;q10) cannot be fully interpreted
due to the absence of the cytogenetic results at
diagnosis. However, concurrence between the
detection of this cytogenetic abnormality and
worsening of the patient's symptoms suggests that
the der(9;18)(p10;q10) may have been an early
marker of the disease evolution.
Gain of 9p resulting from +i(9)(p10) has been
reported in two cases of PV, further indicating this
gain as a recurrent finding in PV. Our patient is
known to carry the activating JAK2 V617F
mutation. One can hypothesize that in combination
with this mutation, gain of an extra copy of either
the mutated or the normal JAK2 allele through
formation of the der(9;18)(p10;q10) contributed to
the progression of the patient's disease. Our report
therefore further suggests the association between
the unbalanced rearrangement der(9;18)(p10;q10)
and an advanced stage of polycythemia vera.
References Chen Z, Notohamiprodjo M, Guan XY, Paietta E, Blackwell S, Stout K, Turner A, Richkind K, Trent JM, Lamb A, Sandberg AA. Gain of 9p in the pathogenesis of polycythemia vera. Genes Chromosomes Cancer. 1998 Aug;22(4):321-4
Bacher U, Haferlach T, Schoch C. Gain of 9p due to an unbalanced rearrangement der(9;18): a recurrent clonal abnormality in chronic myeloproliferative disorders. Cancer Genet Cytogenet. 2005 Jul 15;160(2):179-83
Ohyashiki K, Kodama A, Ohyashiki JH. Recurrent der(9;18) in essential thrombocythemia with JAK2 V617F is highly linked to myelofibrosis development. Cancer Genet Cytogenet. 2008 Oct;186(1):6-11
This article should be referenced as such:
Xu X, Chen X, Rauch EA, Johnson EB, Thompson KJ, Laffin JJS, Raca G, Kurtycz DF. Unbalanced rearrangement der(9;18)(p10;q10) in a patient with polycythemia vera. Atlas Genet Cytogenet Oncol Haematol. 2011; 15(1):114-115.
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