presentation by jennifer unterbrink pcdu seminar november 11th, 2015
TRANSCRIPT
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presentation by Jennifer UnterbrinkPCDU seminar
November 11th, 2015
ESCRT Complex and Retromer
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Endosomes in plants
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Endosomes in plants
www.illuminatedcell.com
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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
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Multivesicular bodies
Winter and Hauser, 2006. Trends in Plant Sc. Vol.11 No.3
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Endosomal trafficking
retrograde transportanterograde transport
biosynthetical/ secretory transportBruce Alberts (Ed.), Molecular biology of the cell
ER
Golgi
EE
secretory vesicles
PM
LE
V/ L
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Coats of the late endosomal system
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Retromer schematic
Schellmann and Pimpl, 2009: Curr. Opinion in Plant Biol. 12
SNX: phox-homology domain, allows association with phospholipids (PI3P)
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BAR domain
Bin-Amphiphysin-Rvs
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Retromer structure
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SNXs important for Retromer function
Rojas et al., 2007. Mol. and Cell. Biol. 02/2007
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Plant Retromer
Schellmann and Pimpl, 2009: Curr. Opinion in Plant Biol. 12
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ESCRTs
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ESCRT subunits in plants
Schellmann and Pimpl, 2009: Curr. Opinion in Plant Biol. 12
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ESCRT subunits in plants
Schellmann and Pimpl, 2009: Curr. Opinion in Plant Biol. 12
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ESCRTs in cytokinesis
Spitzer et al., 2006. Development 133
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The coat to perdition
www.nature.com, 2015/11/08
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ESCRT-dependent protein sorting
Winter and Hauser, 2006. Trends in Plant Sc. 11
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Ubiquitin
www.nobelprize.org/nobel_prizes/chemistry/laureates/2004/chempub1high.jpg
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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.
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FREE1
peripheral membrane-associated protein
Gao et al., 2014. Curr. Biol. 24
binds to PI3P via the FYVE domain
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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
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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
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FREE1 function
Gao et al., 2014. Curr. Biol. 24
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https://www.youtube.com/watch?v=_4DM48ZPR0A