presentation by jennifer unterbrink pcdu seminar november 11th, 2015

26
presentation by Jennifer Unterbrink PCDU seminar November 11 th , 2015 ESCRT Complex and Retromer

Upload: julie-murphy

Post on 19-Jan-2018

214 views

Category:

Documents


0 download

TRANSCRIPT

Page 1: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

presentation by Jennifer UnterbrinkPCDU seminar

November 11th, 2015

ESCRT Complex and Retromer

Page 2: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015
Page 3: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

Endosomes in plants

Page 4: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

Endosomes in plants

www.illuminatedcell.com

Page 5: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

Endosomes in plants heterogenous collection of organelles

function:

sorting and delivery from the cell surface

transport from TGN to lysosome/ vacuole

biosynthetic AND endocytotic pathway

Page 6: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

Multivesicular bodies

Winter and Hauser, 2006. Trends in Plant Sc. Vol.11 No.3

Page 7: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

Endosomal trafficking

retrograde transportanterograde transport

biosynthetical/ secretory transportBruce Alberts (Ed.), Molecular biology of the cell

ER

Golgi

EE

secretory vesicles

PM

LE

V/ L

Page 8: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

Coats of the late endosomal system

Page 9: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

Retromer schematic

Schellmann and Pimpl, 2009: Curr. Opinion in Plant Biol. 12

SNX: phox-homology domain, allows association with phospholipids (PI3P)

Page 10: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

BAR domain

Bin-Amphiphysin-Rvs

Page 11: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

Retromer structure

Page 12: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

SNXs important for Retromer function

Rojas et al., 2007. Mol. and Cell. Biol. 02/2007

Page 13: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

Plant Retromer

Schellmann and Pimpl, 2009: Curr. Opinion in Plant Biol. 12

Page 14: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

ESCRTs

Page 15: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

ESCRT subunits in plants

Schellmann and Pimpl, 2009: Curr. Opinion in Plant Biol. 12

Page 16: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

ESCRT subunits in plants

Schellmann and Pimpl, 2009: Curr. Opinion in Plant Biol. 12

Page 17: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

ESCRTs in cytokinesis

Spitzer et al., 2006. Development 133

Page 18: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

The coat to perdition

www.nature.com, 2015/11/08

Page 19: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

ESCRT-dependent protein sorting

Winter and Hauser, 2006. Trends in Plant Sc. 11

Page 20: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

Ubiquitin

www.nobelprize.org/nobel_prizes/chemistry/laureates/2004/chempub1high.jpg

Page 21: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

ESCRT-dependent vesicle formation

Cai et al., 2014. Plant Physiol. Vol.165 No.3c

Figure 7.Immunogold detection of VSRs in wild-type and transgenic Arabidopsis plants with induced expression of ESCRT-III mutants. Six-day-old wild-type plants (A and B) and transgenic plants (C–J) germinated and cultured in the absence of inducers were transferred onto medium containing 10 μM β-estradiol for 24 h to induce the expression of VPS20.1-1-112-YFP (C and D), SNF7.1(L22W)-YFP (E and F), VPS24.1-1-152-YFP (G and H), or VPS2.1-1-179-YFP (I and J). Arabidopsis roots were subjected to high-pressure frozen/freeze substitution followed by ultrathin sectioning and immunogold labeling using VSR antibodies. Bar = 100 nm.

Page 22: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

FREE1

peripheral membrane-associated protein

Gao et al., 2014. Curr. Biol. 24

binds to PI3P via the FYVE domain

Page 23: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

FYVE domainFYVE zinc finger domainnamed after the Cys-rich proteins Fab1, YOTB, Vac1, EEA1

binds PI3P

is composed of two small β-hairpins (zinc knuckles)followed by a α-helix

FYVE finger binds to two Zn-ions

Page 24: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

FREE1 function

transposon insertion in A.thaliana

Gao et al., 2014. Curr. Biol. 24

homozygous free1 mutants are not viable

expression of GFP-tagged FREE1 rescued the seedling lethality

Page 25: presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

FREE1 function

Gao et al., 2014. Curr. Biol. 24