Very long-chain fatty acid-containing lipids rather than sphingolipids per se are required for raft association and stable surface transport of newly synthesized plasma membrane ATPase in yeast.
The proton-pumping H+-ATPase, Pma1p, is an abundant and very long lived polytopic protein of the yeast plasma membrane. Pma1p constitutes a major cargo of the secretory pathway and thus serves as a model to study plasma membrane biogenesis. Pma1p associates with detergent-resistant membrane domains (lipid "rafts") already in the ER, and a lack of raft association correlates with mistargeting of the protein to the vacuole, where it is degraded. We are analyzing the role of specific lipids in membrane domain formation and have previously shown that surface transport of Pma1p is independent of newly synthesized sterols but that sphingolipids with C26 very long chain fatty acid are crucial for raft association and surface transport of Pma1p (Gaigg, B., Timischl, B., Corbino, L., and Schneiter, R. (2005) J. Biol. Chem. 280, 22515-22522). We now describe a more detailed analysis of the function that sphingolipids play in this process. Using a yeast strain in which the essential function of sphingolipids is substituted by glycerophospholipids containing C26 very long chain fatty acids, we find that sphingolipids per se are dispensable for raft association and surface delivery of Pma1p but that the C26 fatty acid is crucial. We thus conclude that the essential function of sphingolipids for membrane domain formation and stable surface delivery of Pma1p is provided by the C26 fatty acid that forms part of the yeast ceramide.
Gaigg
Barbara
B
Division of Biochemistry, University of Fribourg, CH-1700 Fribourg, Switzerland.
Toulmay
Alexandre
A
Schneiter
Roger
R
D002462:Cell Membrane / Q000201:enzymology*; D004705:Endocytosis; D004795:Enzyme Stability; D005227:Fatty Acids / Q000378:metabolism*; D007527:Isoenzymes; D050356:Lipid Metabolism; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D009154:Mutation; D021381:Protein Transport; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D013107:Sphingolipids / Q000502:physiology*; D013997:Time Factors
D005227:Fatty Acids; D007527:Isoenzymes; D029701:Saccharomyces cerevisiae Proteins; D013107:Sphingolipids; C092477:PMA1 protein, S cerevisiae; D006180:Proton-Translocating ATPases