title: Aureobasidin A arrests growth of yeast cells through both ceramide intoxication and deprivation of essential inositolphosphorylceramides.
abstract: All mature Saccharomyces cerevisiae sphingolipids comprise inositolphosphorylceramides containing C26:0 or C24:0 fatty acids and either phytosphingosine or dihydrosphingosine. Here we analysed the lipid profile of lag1Delta lac1Delta mutants lacking acyl-CoA-dependent ceramide synthesis, which require the reverse ceramidase activity of overexpressed Ydc1p for sphingolipid biosynthesis and viability. These cells, termed 2Delta.YDC1, make sphingolipids containing exclusively dihydrosphingosine and an abnormally wide spectrum of fatty acids with between 18 and 26 carbon atoms. Like wild-type cells, 2Delta.YDC1 cells stop growing when exposed to Aureobasidin A (AbA), an inhibitor of the inositolphosphorylceramide synthase AUR1, yet their ceramide levels remain very low. This finding argues against a current hypothesis saying that yeast cells do not require inositolphosphorylceramides and die in the presence of AbA only because ceramides build up to toxic concentrations. Moreover, W303lag1Delta lac1Delta ypc1Delta ydc1Delta cells, reported to be AbA resistant, stop growing on AbA after a certain number of cell divisions, most likely because AbA blocks the biosynthesis of anomalous inositolphosphorylsphingosides. Thus, data argue that inositolphosphorylceramides of yeast, the equivalent of mammalian sphingomyelins, are essential for growth. Data also clearly confirm that wild-type strains, when exposed to AbA, immediately stop growing because of ceramide intoxication, long before inositolphosphorylceramide levels become subcritical.
authors.affiliation: Department of Medicine, University of Fribourg, Fribourg, Switzerland.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Cerantola
authors.forename: Isabelle
authors.initials: I
authors.lastname: Guillas
authors.forename: Carole
authors.initials: C
authors.lastname: Roubaty
authors.forename: Christine
authors.initials: C
authors.lastname: Vionnet
authors.forename: Danièle
authors.initials: D
authors.lastname: Uldry
authors.forename: Jens
authors.initials: J
authors.lastname: Knudsen
authors.forename: Andreas
authors.initials: A
authors.lastname: Conzelmann
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology; D002518:Ceramides / Q000096:biosynthesis*; D047630:Depsipeptides / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006028:Glycosphingolipids / Q000096:biosynthesis*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism
chemical_list: D000935:Antifungal Agents; D002518:Ceramides; D047630:Depsipeptides; D004791:Enzyme Inhibitors; D006028:Glycosphingolipids; C477937:inositolphosphorylceramide; C071398:aureobasidin A