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presentation by Jennifer Unterbrink PCDU seminar November 11th, 2015

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

2

3 Endosomes in plants

4 Endosomes in plants

5 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

6 Multivesicular bodies
Winter and Hauser, Trends in Plant Sc. Vol.11 No.3

7 Endosomal trafficking
ER Golgi secretory vesicles LE V/ L EE PM retrograde transport anterograde transport biosynthetical/ secretory transport Bruce Alberts (Ed.), Molecular biology of the cell

8 Coats of the late endosomal system

9 Retromer schematic SNX: phox-homology domain, allows association with phospholipids (PI3P) Schellmann and Pimpl, 2009: Curr. Opinion in Plant Biol. 12

10 BAR domain Bin-Amphiphysin-Rvs

11 Retromer structure

12 SNXs important for Retromer function
Rojas et al., Mol. and Cell. Biol. 02/2007

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

14 ESCRTs

15 ESCRT subunits in plants
Schellmann and Pimpl, 2009: Curr. Opinion in Plant Biol. 12

16 ESCRT subunits in plants
Schellmann and Pimpl, 2009: Curr. Opinion in Plant Biol. 12

17 ESCRTs in cytokinesis Spitzer et al., Development 133

18 The coat to perdition 2015/11/08

19 ESCRT-dependent protein sorting
Winter and Hauser, Trends in Plant Sc. 11

20 Ubiquitin

21 ESCRT-dependent vesicle formation
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 VPS YFP (C and D), SNF7.1(L22W)-YFP (E and F), VPS YFP (G and H), or VPS 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. Cai et al., Plant Physiol. Vol.165 No.3c

22 FREE1 peripheral membrane-associated protein
binds to PI3P via the FYVE domain Gao et al., Curr. Biol. 24

23 FYVE domain FYVE zinc finger domain
named after the Cys-rich proteins Fab1, YOTB, Vac1, EEA1 is composed of two small β-hairpins (zinc knuckles) followed by a α-helix binds PI3P FYVE finger binds to two Zn-ions

24 FREE1 function transposon insertion in A.thaliana
homozygous free1 mutants are not viable expression of GFP-tagged FREE1 rescued the seedling lethality Gao et al., Curr. Biol. 24

25 FREE1 function Gao et al., Curr. Biol. 24

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