title: A novel RING finger protein complex essential for a late step in protein transport to the yeast vacuole.
abstract: Protein transport to the lysosome-like vacuole in yeast is mediated by multiple pathways, including the biosynthetic routes for vacuolar hydrolases, the endocytic pathway, and autophagy. Among the more than 40 genes required for vacuolar protein sorting (VPS) in Saccharomyces cerevisiae, mutations in the four class C VPS genes result in the most severe vacuolar protein sorting and morphology defects. Herein, we provide complementary genetic and biochemical evidence that the class C VPS gene products (Vps18p, Vps11p, Vps16p, and Vps33p) physically and functionally interact to mediate a late step in protein transport to the vacuole. Chemical cross-linking experiments demonstrated that Vps11p and Vps18p, which both contain RING finger zinc-binding domains, are components of a hetero-oligomeric protein complex that includes Vps16p and the Sec1p homologue Vps33p. The class C Vps protein complex colocalized with vacuolar membranes and a distinct dense membrane fraction. Analysis of cells harboring a temperature-conditional vps18 allele (vps18tsf) indicated that Vps18p function is required for the biosynthetic, endocytic, and autophagic protein transport pathways to the vacuole. In addition, vps18tsf cells accumulated multivesicular bodies, autophagosomes, and other membrane compartments that appear to represent blocked transport intermediates. Overproduction of either Vps16p or the vacuolar syntaxin homologue Vam3p suppressed defects associated with vps18tsf mutant cells, indicating that the class C Vps proteins and Vam3p may functionally interact. Thus we propose that the class C Vps proteins are components of a hetero-oligomeric protein complex that mediates the delivery of multiple transport intermediates to the vacuole.
authors.affiliation: Department of Biology, Howard Hughes Medical Institute, University of California, San Diego, School of Medicine, La Jolla 92093-0668, USA.
authors.forename: S E
authors.initials: SE
authors.lastname: Rieder
authors.forename: S D
authors.initials: SD
authors.lastname: Emr
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport; D000483:Alleles; D001343:Autophagy / Q000502:physiology; D001692:Biological Transport; D002458:Cell Fractionation; D004705:Endocytosis / Q000502:physiology; D005656:Fungal Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D007425:Intracellular Membranes / Q000737:chemistry; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D029701:Saccharomyces cerevisiae Proteins; D013696:Temperature; D014617:Vacuoles / Q000201:enzymology / Q000378:metabolism* / Q000648:ultrastructure; D033921:Vesicular Transport Proteins; D016335:Zinc Fingers
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D005656:Fungal Proteins; D008565:Membrane Proteins; C071657:PEP3 protein, S cerevisiae; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; C080045:VPS16 protein, S cerevisiae; D033921:Vesicular Transport Proteins