Proteomic and Biochemical Comparison of the Cellular Interaction Partners of Human VPS33A and VPS33B

Journal of Molecular Biology
Morag R HunterStephen C Graham

Abstract

Multi-subunit tethering complexes control membrane fusion events in eukaryotic cells. Class C core vacuole/endosome tethering (CORVET) and homotypic fusion and vacuole protein sorting (HOPS) are two such complexes, both containing the Sec1/Munc18 protein subunit VPS33A. Metazoans additionally possess VPS33B, which has considerable sequence similarity to VPS33A but does not integrate into CORVET or HOPS complexes and instead stably interacts with VIPAR. It has been recently suggested that VPS33B and VIPAR comprise two subunits of a novel multi-subunit tethering complex (named "CHEVI"), perhaps analogous in configuration to CORVET and HOPS. We utilized the BioID proximity biotinylation assay to compare and contrast the interactomes of VPS33A and VPS33B. Overall, few proteins were identified as associating with both VPS33A and VPS33B, suggesting that these proteins have distinct sub-cellular localizations. Consistent with previous reports, we observed that VPS33A was co-localized with many components of class III phosphatidylinositol 3-kinase (PI3KC3) complexes: PIK3C3, PIK3R4, NRBF2, UVRAG and RUBICON. Although VPS33A clearly co-localized with several subunits of CORVET and HOPS in this assay, no proteins with the canonical CORVE...Continue Reading

Citations

May 10, 2019·Human Molecular Genetics·Elena V PavlovaTimothy M Cox
Jan 23, 2020·Journal of Cell Science·Daniela SparvoliAaron P Turkewitz
May 17, 2019·Journal of Cell Science·Jan van der BeekJudith Klumperman
Oct 16, 2020·Cellular and Molecular Life Sciences : CMLS·Anna CiesielskaKatarzyna Kwiatkowska

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Datasets Mentioned

BETA
MSV000081814

Methods Mentioned

BETA
GTPases
immunoprecipitation
co-immunoprecipitation
size exclusion chromatography
pull-down
gel filtration
PCR
affinity purification
immunoprecipitations

Software Mentioned

CORVET
ProHits LIMS
GeneArt
BioID
TASSER
WASH
Retriever
SAINT
Image Studio Lite
Retromer

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