the small ubiquitin-like modifiers: established and emerging roles in diseases mike tatham

The Small Ubiquitin-like Modifiers: Established and emerging roles in diseases

Mike Tatham

Ron Hay labWellcome Trust Centre for Gene Regulation and Expression

University of Dundee

ELRIG/SLAS Drug Discovery Manchester 2012


Phylogenetic relationship in the ubiquitin like modifier superfamily

• SUMO system is only found in Eukaryotes• Yeasts, flies and worms only express a single SUMO • Vertebrates express three paralogues SUMO-1, SUMO-

2, SUMO-3


The SUMO conjugation system

Ubiquitination

Substrate

Ubiquitin E1 (2)Ubiquitin E2 (~20)Ubiquitin E3 (hundreds) UU

UU

SubstrateUbiquitin

Protease (~100)

SS

S S

S

SUMOylation

SUMO E1 (1 – SAE1/2)SUMO E2 (1 – Ubc9)[SUMO E3 (10-20?)]

SUMOProtease (8)

SubstrateSubstrate


2, SUMO-3 • Like ubiquitin SUMOs are conjugated to protein

substrates in a three step mechanism• Most SUMO conjugation occurs within a consensus

motif yKXE/D• SUMO conjugation can occur independent of E3s• SUMO-2 and SUMO-3 contain consensus motifs and

can modify themselves to form polySUMO chains• Deletion of yeast SUMO is lethal• Deletion of Ubc9 in mice is lethal

yKXE yKXD

Structural overview





can modify themselves to form polySUMO chains• Deletion of yeast SUMO is lethal• Deletion of Ubc9 in mice is lethal• SUMOs have low sequence homology to ubiquitin but

high 3D structural similarity


Cellular characteristics

IB Ubiquitin

IB SUMO-2/3

IB SUMO-1

UU

UU

Substrate

U

SS

SS

Substrate

S

Substrate

S

S

Time (h)

+MG132

0 1 2 3 5 7

Total extracts






high 3D structural similarity• SUMOs are predominantly nuclear proteins• SUMO-1 and SUMO-2/-3 have largely overlapping

protein targets with some distinctions


Molecular functions of SUMO

SS

S S

S

SUMOylation

SUMO E1 (1 – SAE1/2)SUMO E2 (1 – Ubc9)[SUMO E3 (10-20?)]

SUMOProtease (8)

SubstrateSubstrate

SUMO BP

SUMO BP

polySUMO BP

Substrate

S

SSubstrate

S

Substrate

SS

S S

Subcellular localisationEnzymatic activity

Complex formationFurther modificationBlock modifications

Altered function






high 3D structural similarity• SUMOs are predominantly nuclear proteins• SUMO-1 and SUMO-2/-3 have largely overlapping

protein targets with some distinctions• SUMO conjugation does not have a common effect on

proteins, but has myriad of protein-specific consequences mediated by SUMO Interaction Motifs (SIMS)

Cellular functions of SUMO

TAP-SUMO-2 cells

•Purify SUMO from cells•Identify and quantify proteins by quantitative mass spectrometry-based proteomics•Identified a total of ~900 SUMO substrates

Num

ber o

f SU

MO

sub

stra

tes

Year

SubstrateTAG SUMO


~10% of cellular proteins are modified by SUMO

Golebiowski et al Sci Signal. 2009;2(72):ra24. Tatham et al Sci. Signal. 2011;4(178):rs4

SUMO


SUMO and human diseases

Immunologicaldisorders

Centromere instability, and facial anomalies syndrome

Autoimmuneregulation

Dermatomyositis

Infectiousdiseases

DNA viruses Protozoa

Extra and Intra-cellular bacteria

RNA viruses

CirculatoryDiseases

Transient global and focal cerebral ischemia

Familial dilated cardiomyopathy

Congenital heart disease

Obesity

Others

Rheumatoid arthritis

Liver damage

Cystic fibrosis

Musculardystrophy

Heart failure

Neurologicaldisorders

Multiple System Atrophy

Alzheimer's disease

Frontotemporal dementia

Spinal and bulbar muscular atrophy

Huntington's disease

Amyotrophic lateral sclerosis

Frontotemporal lobar degeneration

Spinocerebellar ataxia type 1

Neuronal intranuclear inclusion disease Dementia with Lewy Bodies

Parkinson's disease


Cancers

Megakaryoblastic leukemia

Squamous cell carcinoma

Colon cancer

Acute Promyelocytic leukaemia

Breast cancer

Ovarian cancer Multiple myeloma

MelanomaProstate cancer

Atypical myeloproliferative disease

Renal cell carcinoma



Types of evidence linking SUMO with diseases1. Disease protein x is modified by SUMO which alters its function.2. SUMO conjugation is altered in disease cells3. SUMO is abnormally distributed within disease cells4. Enzymes of the SUMO modification system are abnormally expressed in disease cells5. SUMO system modulation alters the disease phenotype in model cells

Alzheimer’s disease (Amyloid beta protein (derived from from APP))(tau)

Parkinson’s disease (a-synuclein, DJ-1)Prion disease (PrP)Polyglutamine diseases Huntington’s (Huntingtin)

Kennedy’s (Androgen Rec)Dentatorubro-pallidoluysian atrophy (Atrophin-1)Spinocerebellar ataxia (ATAXIN1, 7)Tauopathy (tau)Familial amyotrophic lateral sclerosis (SOD1) Fei et al. BBRC 347 (2006)

Riley et al. JBC 280 (2005), Janer et al. Hum. Mol. Gen. 19 (2010)

Li et al. PNAS 100 (2003)

Juanes et al. JBC 284 (2009)

Mukherjee et al. JBC (2009)Steffan et al. Science (2004)

Dorval & Fraser. JBC 281 (2005), Shinbo et al. Cell. Death Diff. (2006)Dorval & Fraser. JBC 281 (2005)

Dorval & Fraser. JBC 281 (2005)

Terashima et al. Neuroreport. 13 (2002)

Immunolabelling of NII in the hippocampal subiculum of patients with NIID. (Takahashi-Fujigasaki et al. Neuropathology and Applied Neurobiology (2006), 32 , 92–100)

Immunostaining of aggregates in glioma cell models for multiple system atrophy (MSA).

(D.L. Pountney et al. Neuroscience Letters 381 (2005) 74–79)

SUMO BP

SUMO BP


The SUMO-SIM interaction

SUMO

SIM peptide

Q. How do we take advantage of the SUMO system therapeutically?A. It depends on what you want to do!

SS

S S

SSUMO E1 (1 – SAE1/2)SUMO E2 (1 – Ubc9)[SUMO E3 (10-20?)]

SUMOProtease (8)

SubstrateSubstrateyKXE yKXD

Wimmer P, et al. J Virol. 2012 Jan;86(2):642-54.

To be or not to be specific: What can parasites tell us?

Infectiousdiseases



RNA viruses


Q. How do we take advantage of the SUMO system therapeutically?A. It depends on how specific you want to be!

SUMO



Immunologicaldisorders

Centromere instability, and facial anomalies syndrome

Autoimmuneregulation

Dermatomyositis

Infectiousdiseases



RNA viruses

CirculatoryDiseases

Transient global and focal cerebral ischemia

Familial dilated cardiomyopathy

Congenital heart disease

Obesity

Others

Rheumatoid arthritis

Liver damage

Cystic fibrosis

Musculardystrophy

Heart failure

Neurologicaldisorders

Multiple System Atrophy

Alzheimer's disease

Frontotemporal dementia

Spinal and bulbar muscular atrophy

Huntington's disease


Frontotemporal lobar degeneration

Spinocerebellar ataxia type 1

Neuronal intranuclear inclusion disease Dementia with Lewy Bodies

Parkinson's disease


Cancers

Megakaryoblastic leukemia

Squamous cell carcinoma

Colon cancer

Acute Promyelocytic leukaemia

Breast cancer

Ovarian cancer Multiple myeloma

MelanomaProstate cancer

Atypical myeloproliferative disease

Renal cell carcinoma


Example of a successful drug therapy involving SUMO - APL

De The et al J. Cell. Biol. 2012. 198 No.1 11-21 Liu et al Curr. Op. Chem. Biol. 2012. 16 92-98

Acute Promyelocytic Leukaemia (APL)

•A rare condition driven by a chromosomal translocation resulting in the fusion of the PML and retinoic acid receptor a proteins (PML-RARa)

•Very malignant and charaterised by sudden hemorrhages and accumulation of promyelocytes in blood

•Retinoic acid and arsenic trioxide treatment induce differentiation of promyelocytes and clinical remission.

•PML-RARa and PML are known to be SUMOylated and degraded in response to arsenic



Ring Finger protein 4 (RNF4) aka SNURF

Tatham et al 2008. Nat. Cell. Biol. 10. 5. 538-546


A model for SUMO-dependent disease remission

PML

SUMOconjugation

SUMOdeconjugation

Ubiquitinconjugation

Ubiquitindeconjugation

U

PML

SS

S S S

PML

SS

S S SU

UUU

UUU

U

ARSENIC RNF4



The SUMO system as a therapeutic target - Summary

•SUMO is functionally highly pleiotropic affecting many important cellular pathways

•There is a range of evidence linking SUMO to significant human diseases.

•The precise role of SUMO in many diseases is not determined and so its potential as a therapeutic target is largely unclear

•The best approach to modulating SUMO function for individual disease therapy is unclear

•There is an academic and clinical argument for small molecule effectors of the SUMO system to help clarify these issues.

Ron HayFilip Golebiowski (Glasgow)Ellis JaffrayMarie-Claude Geoffroy (Paris)Ivan MaticAmit Garg

Jurgen CoxMatthias Mann


Acknowledgements

the small ubiquitin-like modifiers: established and emerging roles in diseases mike tatham

Documents

paralogues sumo

single sumo vertebrates

ubiquitin e2

ubiquitin sumos

small ubiquitin

consensus motifs

substratesubstratesumo

protein substrates