title: Actin polymerization driven by WASH causes V-ATPase retrieval and vesicle neutralization before exocytosis.
abstract: WASP and SCAR homologue (WASH) is a recently identified and evolutionarily conserved regulator of actin polymerization. In this paper, we show that WASH coats mature Dictyostelium discoideum lysosomes and is essential for exocytosis of indigestible material. A related process, the expulsion of the lethal endosomal pathogen Cryptococcus neoformans from mammalian macrophages, also uses WASH-coated vesicles, and cells expressing dominant negative WASH mutants inefficiently expel C. neoformans. D. discoideum WASH causes filamentous actin (F-actin) patches to form on lysosomes, leading to the removal of vacuolar adenosine triphosphatase (V-ATPase) and the neutralization of lysosomes to form postlysosomes. Without WASH, no patches or coats are formed, neutral postlysosomes are not seen, and indigestible material such as dextran is not exocytosed. Similar results occur when actin polymerization is blocked with latrunculin. V-ATPases are known to bind avidly to F-actin. Our data imply a new mechanism, actin-mediated sorting, in which WASH and the Arp2/3 complex polymerize actin on vesicles to drive the separation and recycling of proteins such as the V-ATPase.
authors: Cancer Research UK Beatson Institute for Cancer Research, Bearsden, Glasgow G61 1BD, Scotland, UK.
authors: Michael
authors: M
authors: Carnell
authors: Tobias
authors: T
authors: Zech
authors: Simon D
authors: SD
authors: Calaminus
authors: Seiji
authors: S
authors: Ura
authors: Monica
authors: M
authors: Hagedorn
authors: Simon A
authors: SA
authors: Johnston
authors: Robin C
authors: RC
authors: May
authors: Thierry
authors: T
authors: Soldati
authors: Laura M
authors: LM
authors: Machesky
authors: Robert H
authors: RH
authors: Insall
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