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Yeast sphingolipids do not need to contain very long chain fatty acids

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  • Biochemical Journal. - 2007, vol. 401, no. 1, p. 205-216
English Synthesis of VLCFAs (very long chain fatty acids) and biosynthesis of DHS (dihydrosphingosine) both are of vital importance for Saccharomyces cerevisiae. The bulk of VLCFAs and DHS are used for ceramide synthesis by the Lag1p (longevity-assurance gene 1)/Lac1p (longevity-assurance gene cognate 1)/Lip1p (Lag1p/Lac1p interacting protein) ceramide synthase. LAG1 and LAC1 are redundant but LIP1 is essential. Here we show that 4D (lag1Dlac1Dypc1Dydc1D) cells devoid of all known endogenous ceramide synthesis pathways are unviable but can be rescued by the expression of Lass5, a mouse LAG1 homologue. Ceramide synthase activity of 4D.Lass5 cells only utilizes C₁₆ and C₁₈ fatty acids and does not require the help of Lip1p, an essential cofactor of Lag1p/Lac1p. HPLC-electrospray ionization-MS/MS analysis demonstrated that in IPCs (inositolphosphorylceramides) of 4D.Lass5, the very long chain fatty acids (C₂₆ and C₂₄) account for <1% instead of the normal >97%. Notwithstanding, IPCs incorporated into glycosylphosphatidylinositol anchors of 4D.Lass5 show normal mobility on TLC and the ceramide- and raft-dependent traffic of Gas1p (glycophospholipid-anchored surface protein) from endoplasmic reticulum to Golgi remains almost normal. Moreover, the biosynthesis of C24:0 fatty acids remains essential. Thus, C24:0 and dihydrosphingosine are both necessary for survival of yeast cells even if they utilize C₁₆ and C₁₈ fatty acids for sphingolipid biosynthesis.
Faculté des sciences et de médecine
Département de Biologie, Département de Chimie
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