Document Detail


Endosomal recruitment of the WASH complex: active sequences and mutations impairing interaction with the retromer.
MedLine Citation:
PMID:  23331060     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND INFORMATION: The Wiskott-Aldrich syndrome protein and scar homolog (WASH) complex is the major Arp2/3 activator at the surface of endosomes. The branched actin network, that the WASH complex induces, contributes to cargo sorting and scission of transport intermediates destined for most endosomal routes. A major challenge is to understand how the WASH molecular machine is recruited to the surface of endosomes. The retromer endosomal machinery has been proposed by us and others to play a role in this process.
RESULTS: In this work, we used an unbiased approach to identify the endosomal receptor of the WASH complex. We have delineated a short fragment of the FAM21 subunit that is able to displace the endogenous WASH complex from endosomes. Using a proteomic approach, we have identified the retromer cargo selective complex (CSC) as a partner of the active FAM21 sequence displacing the endogenous WASH complex. A point mutation in FAM21 that abolishes CSC interaction also impairs WASH complex displacement activity. The CSC is composed of three subunits, VPS35, VPS29 and VPS26. FAM21 directly binds the VPS35 subunit of the retromer CSC. Additionally, we show that a point mutant of VPS35 that blocks binding to VPS29 also prevents association with FAM21 and the WASH complex revealing a novel role for the VPS35-VPS29 interaction in regulating retromer association with the WASH complex.
CONCLUSIONS: This novel approach of endogenous WASH displacement confirms previous suggestions that the retromer is the receptor of the WASH complex at the surface of endosomes and identify key residues that mediate this interaction. The interaction between these two endosomal machineries, the WASH complex and the retromer, is likely to play a critical role in forming platforms at the surface of endosomes for efficient sorting of cargoes.
Authors:
Emmanuèle Helfer; Michael E Harbour; Véronique Henriot; Goran Lakisic; Carla Sousa-Blin; Larisa Volceanov; Matthew N J Seaman; Alexis Gautreau
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2013-03-07
Journal Detail:
Title:  Biology of the cell / under the auspices of the European Cell Biology Organization     Volume:  105     ISSN:  1768-322X     ISO Abbreviation:  Biol. Cell     Publication Date:  2013 May 
Date Detail:
Created Date:  2013-05-03     Completed Date:  2013-11-11     Revised Date:  2014-03-14    
Medline Journal Info:
Nlm Unique ID:  8108529     Medline TA:  Biol Cell     Country:  England    
Other Details:
Languages:  eng     Pagination:  191-207     Citation Subset:  IM    
Copyright Information:
© 2013 Société Française des Microscopies and Société de Biologie Cellulaire de France.
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MeSH Terms
Descriptor/Qualifier:
Animals
Binding Sites
Binding, Competitive
Carrier Proteins / chemistry,  genetics,  metabolism*
Endosomes / metabolism*
HeLa Cells
Humans
Mice
Microfilament Proteins / metabolism*
Models, Molecular
Multiprotein Complexes / chemistry,  genetics,  metabolism
Mutation, Missense
NIH 3T3 Cells
Point Mutation
Protein Binding
Protein Structure, Secondary
Protein Structure, Tertiary
Protein Subunits / chemistry,  genetics,  metabolism
Vesicular Transport Proteins / chemistry,  genetics,  metabolism*
Grant Support
ID/Acronym/Agency:
100140//Wellcome Trust; G0701444//Medical Research Council
Chemical
Reg. No./Substance:
0/Carrier Proteins; 0/FAM21 protein, mouse; 0/Microfilament Proteins; 0/Multiprotein Complexes; 0/Protein Subunits; 0/VPS29 protein, mouse; 0/Vesicular Transport Proteins; 0/Vps26 protein, mouse; 0/Vps35 protein, mouse; 0/WASH protein, mouse

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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