Lag1p and Lac1p Are Essential for the Acyl-CoA–dependent Ceramide Synthase Reaction in Saccharomyces cerevisae

Abstract

Lag1p and Lac1p are two homologous transmembrane proteins of the endoplasmic reticulum in Saccharomyces cerevisiae.Homologous genes have been found in a wide variety of eukaryotes. In yeast, both genes, LAC1 and LAG1, are required for efficient endoplasmic reticulum-to-Golgi transport of glycosylphosphatidylinositol-anchored proteins. In this study, we show that lag1Δlac1Δ cells have reduced sphingolipid levels due to a block of the fumonisin B1-sensitive and acyl-CoA–dependent ceramide synthase reaction. The sphingolipid synthesis defect inlag1Δlac1Δ cells can be partially corrected by overexpression of YPC1 orYDC1, encoding ceramidases that have been reported to have acyl-CoA–independent ceramide synthesis activity. Quadruple mutant cells (lag1Δlac1Δypc1Δydc1Δ) do not make any sphingolipids, but are still viable probably because they produce novel lipids. Moreover,lag1Δlac1Δ cells are resistant to aureobasidin A, an inhibitor of the inositolphosphorylceramide synthase, suggesting that aureobasidin A may be toxic because it leads to increased ceramide levels. Based on these data, LAG1and LAC1 are the first genes to be identified that are required for the fumonisin B1-sensitive and acyl-CoA–dependent ceramide synthase reaction.