Role for Drs2p, a P-type ATPase and potential aminophospholipid translocase, in yeast late Golgi function.
ADP-ribosylation factor appears to regulate the budding of both COPI and clathrin-coated transport vesicles from Golgi membranes. An arf1Delta 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 drs2Delta 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-alpha-factor and the accumulation of abnormal Golgi cisternae. Moreover, we observed a marked reduction in clathrin-coated vesicles that can be isolated from the drs2Delta 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.
Chen
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Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37235, USA.
Ingram
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Rosal
P H
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Graham
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TR
D020823:ADP-Ribosylation Factor 1 / Q000235:genetics / Q000502:physiology; D000595:Amino Acid Sequence; D001224:Aspartic Acid / Q000235:genetics / Q000378:metabolism; D001692:Biological Transport; D000252:Calcium-Transporting ATPases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002966:Clathrin / Q000235:genetics / Q000502:physiology; D033922:Clathrin Heavy Chains; D018699:Coated Vesicles / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005800:Genes, Fungal / Q000235:genetics / Q000502:physiology; D005804:Genes, Lethal / Q000235:genetics; D006056:Golgi Apparatus / Q000201:enzymology / Q000378:metabolism*; D007425:Intracellular Membranes / Q000201:enzymology / Q000378:metabolism; D008364:Mannosyltransferases; D000072235:Mating Factor; D008562:Membrane Glycoproteins / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D015388:Organelles / Q000378:metabolism; D010455:Peptides / Q000378:metabolism; D010641:Phenotype; D049294:Phospholipid Transfer Proteins; D043484:Proprotein Convertases; D011498:Protein Precursors / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D013381:Subtilisins / Q000378:metabolism
D002352:Carrier Proteins; D002966:Clathrin; C104014:DRS2 protein, S cerevisiae; D005656:Fungal Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D010455:Peptides; D049294:Phospholipid Transfer Proteins; D011498:Protein Precursors; D029701:Saccharomyces cerevisiae Proteins; D033922:Clathrin Heavy Chains; D001224:Aspartic Acid; D000072235:Mating Factor; C086604:MNN1 protein, S cerevisiae; D008364:Mannosyltransferases; D043484:Proprotein Convertases; D013381:Subtilisins; C058751:KEX2 protein, S cerevisiae; D020823:ADP-Ribosylation Factor 1; D000252:Calcium-Transporting ATPases