Role for Drs2p, a P-Type ATPase and Potential Aminophospholipid Translocase, in Yeast Late Golgi Function

• Published in: The Journal of cell biology Vol. 147; no. 6; pp. 1223 - 1236
• Main Authors: Chen, Chih-Ying, Ingram, Michael F., Rosal, Peter H., Graham, Todd R.
• Format: Journal Article
• Language: English
• Published: United States Rockefeller University Press 13.12.1999; The Rockefeller University Press
• Subjects: Adenosine triphosphatases; ADP-Ribosylation Factor 1 - genetics; ADP-Ribosylation Factor 1 - physiology; Alleles; Amino Acid Sequence; Aminophospholipid translocase; Antibodies; Aspartic Acid - genetics; Aspartic Acid - metabolism; Biological Transport; Calcium-Transporting ATPases - chemistry; Calcium-Transporting ATPases - genetics; Calcium-Transporting ATPases - metabolism; Carrier Proteins - chemistry; Carrier Proteins - genetics; Carrier Proteins - metabolism; Cell membranes; Cells; chc1 gene; Clathrin - genetics; Clathrin - physiology; Clathrin Heavy Chains; Coated Vesicles - metabolism; COPI protein; Drs2 protein; Drs2p protein; Endosomes - metabolism; Enzymes; Fungal Proteins - chemistry; Fungal Proteins - genetics; Fungal Proteins - metabolism; Genes, Fungal - genetics; Genes, Fungal - physiology; Genes, Lethal - genetics; Golgi Apparatus - enzymology; Golgi Apparatus - metabolism; Intracellular Membranes - enzymology; Intracellular Membranes - metabolism; Kex2 protein; Lipids; Mannosyltransferases; Mating Factor; Membrane Glycoproteins - metabolism; Membrane Proteins - chemistry; Membrane Proteins - genetics; Membrane Proteins - metabolism; Membranes; Molecular biology; Molecular Sequence Data; Mutation; Mutation - genetics; Organelles - metabolism; Original; P branes; Peptides - metabolism; Phenotype; Phospholipid Transfer Proteins; Plasmids; Proprotein Convertases; Protein Precursors - metabolism; Proteins; Saccharomyces cerevisiae; Saccharomyces cerevisiae - cytology; Saccharomyces cerevisiae - enzymology; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins; SEC gene; Subtilisins - metabolism; SWA3/DRS2 gene; Vacuoles; Yeast; Yeasts
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.147.6.1223

ADP-ribosylation factor appears to regulate the budding of both COPI and clathrin-coated transport vesicles from Golgi membranes. An arf1Δ synthetic lethal screen identified SWA3/DRS2, which encodes an integral membrane P-type ATPase and potential aminophospholipid translocase (or flippase). The drs2 null allele is also synthetically lethal with clathrin heavy chain (chc1) temperature-sensitive alleles, but not with mutations in COPI subunits or other SEC genes tested. Consistent with these genetic analyses, we found that the drs2Δ mutant exhibits late Golgi defects that may result from a loss of clathrin function at this compartment. These include a defect in the Kex2-dependent processing of pro-α-factor and the accumulation of abnormal Golgi cisternae. Moreover, we observed a marked reduction in clathrin-coated vesicles that can be isolated from the drs2Δ cells. Subcellular fractionation and immunofluorescence analysis indicate that Drs2p localizes to late Golgi membranes containing Kex2p. These observations indicate a novel role for a P-type ATPase in late Golgi function and suggest a possible link between membrane asymmetry and clathrin function at the Golgi complex.