Drs2p-dependent formation of exocytic clathrin-coated vesicles in vivo
| Autor: | Walter E. Gall, Zhaolin Hua, Nathan C Geething, Sophie I Chen, Ke Liu, Michael F. Ingram, Todd R. Graham |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
Saccharomyces cerevisiae Proteins
Endosome Adaptor Protein Complex 1 Genes Fungal Vesicular Transport Proteins Calcium-Transporting ATPases Saccharomyces cerevisiae Clathrin General Biochemistry Genetics and Molecular Biology Exocytosis 03 medical and health sciences symbols.namesake Translocase Integral membrane protein 030304 developmental biology 0303 health sciences biology Agricultural and Biological Sciences(all) Biochemistry Genetics and Molecular Biology(all) Vesicle 030302 biochemistry & molecular biology Clathrin-Coated Vesicles Golgi apparatus Actin cytoskeleton Phosphoproteins Cell biology Microscopy Electron Clathrin Heavy Chains Mutation symbols biology.protein Clathrin adaptor proteins General Agricultural and Biological Sciences Carrier Proteins Gene Deletion |
| Zdroj: | Current biology : CB. 12(18) |
| ISSN: | 0960-9822 |
| Popis: | The small GTP binding protein ARF has been implicated in budding clathrin-coated vesicles (CCVs) from Golgi and endosomal membranes. An arf1 synthetic lethal screen identified DRS2/SWA3 along with a clathrin heavy-chain conditional allele ( chc1-5/swa5-1 ) and SWA2 , encoding the yeast auxilin-like protein involved in uncoating CCVs [1–3]. Drs2p/Swa3p is a P-type ATPase and a potential aminophospholipid translocase that localizes to the trans -Golgi network (TGN) in yeast. Genetic and phenotypic analyses of drs2Δ mutants suggested that Drs2p was required for clathrin function [1, 4]. To address a potential role for Drs2p in CCV formation from the TGN in vivo, we have performed epistasis analyses between drs2 and mutations that cause accumulation of distinct populations of post-Golgi vesicles. We find that Drs2p is required to form a specific class of secretory vesicles that accumulate when the actin cytoskeleton is disrupted. Accumulation of these vesicles also requires clathrin and is perturbed by mutation of AP-1, but not AP-2, AP-3, or GGA adaptins. Most of the accumulated vesicles are uncoated; however, clathrin coats can be partially stabilized on these vesicles by deletion of SWA2 . These data provide the first in vivo evidence for an integral membrane protein requirement in forming CCVs. |
| Databáze: | OpenAIRE |
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