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PMID: 25783203 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Recruitment of VPS33A to HOPS by VPS16 Is Required for Lysosome Fusion with Endosomes and Autophagosomes.

Traffic (Copenhagen, Denmark) ·Vol. 16 ·No. 7 ·2015-07-00 ·Pages 727-42

Wartosch L, Günesdogan U, Graham SC, Luzio JP

Abstract

The mammalian homotypic fusion and vacuole protein sorting (HOPS) complex is comprised of six subunits: VPS11, VPS16, VPS18, VPS39, VPS41 and the Sec1/Munc18 (SM) family member VPS33A. Human HOPS has been predicted to be a tethering complex required for fusion of intracellular compartments with lysosomes, but it remains unclear whether all HOPS subunits are required. We showed that the whole HOPS complex is required for fusion of endosomes with lysosomes by monitoring the delivery of endocytosed fluorescent dextran to lysosomes in cells depleted of individual HOPS proteins. We used the crystal structure of the VPS16/VPS33A complex to design VPS16 and VPS33A mutants that no longer bind each other and showed that, unlike the wild-type proteins, these mutants no longer rescue lysosome fusion with endosomes or autophagosomes in cells depleted of the endogenous proteins. There was no effect of depleting either VIPAR or VPS33B, paralogs of VPS16 and VPS33A, on fusion of lysosomes with either endosomes or autophagosomes and immunoprecipitation showed that they form a complex distinct from HOPS. Our data demonstrate the necessity of recruiting the SM protein VPS33A to HOPS via its interaction with VPS16 and that HOPS proteins, but not VIPAR or VPS33B, are essential for fusion of endosomes or autophagosomes with lysosomes.

Keywords
CORVET HOPS SM protein VPS16 VPS33A autophagy endocytosis lysosomes tethering factor
MeSH Terms
Amino Acid Sequence Autophagy Endosomes/metabolism HeLa Cells Humans Lysosomes/metabolism Molecular Sequence Data Phagosomes/metabolism Protein Binding Protein Transport Vesicular Transport Proteins/chemistry,genetics,metabolism
Chemicals
VPS16 protein, human VPS33A protein, human Vesicular Transport Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Wartosch Lena
Cambridge Institute for Medical Research and Department of Clinical Biochemistry, Wellcome Trust/MRC Building, University of Cambridge, Cambridge, CB2 0XY, UK.
Günesdogan Ufuk
Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge, CB2 1QN, UK.
Graham Stephen C
Department of Pathology, University of Cambridge, Cambridge, CB2 1QP, UK.
Luzio J Paul
Cambridge Institute for Medical Research and Department of Clinical Biochemistry, Wellcome Trust/MRC Building, University of Cambridge, Cambridge, CB2 0XY, UK.
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Article Info
Journal
Traffic (Copenhagen, Denmark)
Abbr.
Traffic
ISSN
1600-0854
DOI
10.1111/tra.12283
Published
2015-07-00
Epub
2015-00-30
Pages
727-42
Language
English
Region
England
NLM ID
100939340
PMCID
PMC4510706
Subset
IM
Grants
Wellcome Trust · 093026 · United Kingdom
Wellcome Trust · 098406/Z/12/Z · United Kingdom
Wellcome Trust · 079919 · United Kingdom
Wellcome Trust · 098406 · United Kingdom
Medical Research Council · G0900113 · United Kingdom
Wellcome Trust · 100140 · United Kingdom
External Links
PubMed source DOI: 10.1111/tra.12283 PMC Full Text View journal details
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