title: Identification of six complementation classes involved in the biosynthesis of glycosylphosphatidylinositol anchors in Saccharomyces cerevisiae.
abstract: Glycosylphosphatidylinositol (GPI)-anchored membrane proteins are synthesized by the posttranslational attachment of a preformed glycolipid to newly made glycoproteins. alpha-Agglutinin is a GPI-anchored glycoprotein that gets expressed at the cell surface of MAT alpha cells after induction with type a mating factor. Mutants affecting the biosynthesis of GPI anchors were obtained by selecting for the absence of alpha-agglutinin from the cell wall after induction with a-factor at 37 degrees C. 10 recessive mutants were grouped into 6 complementation classes, gpi4 to gpi9. Mutants are considered to be deficient in the biosynthesis of GPI anchors, since each mutant accumulates an abnormal, incomplete GPI glycolipid containing either zero, two, or four mannoses. One mutant accumulates a complete precursor glycolipid, suggesting that it might be deficient in the transfer of complete precursor lipids to proteins. When labeled with [2-3H]inositol, mutants accumulate reduced amounts of radiolabeled GPI-anchored proteins, and the export of the GPI-anchored Gas1p out of the ER is severely delayed in several mutant strains. On the other hand, invertase and acid phosphatase are secreted by all but one mutant. All mutants show an increased sensitivity to calcofluor white and hygromycin B. This suggests that GPI-anchored proteins are required for the integrity of the yeast cell wall.
authors.affiliation: Institute of Biochemistry, University of Fribourg, Switzerland.
authors.forename: M
authors.initials: M
authors.lastname: Benghezal
authors.forename: P N
authors.initials: PN
authors.lastname: Lipke
authors.forename: A
authors.initials: A
authors.lastname: Conzelmann
mesh_terms: D006031:Glycosylation; D017261:Glycosylphosphatidylinositols / Q000096:biosynthesis* / Q000235:genetics; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D013696:Temperature
chemical_list: D017261:Glycosylphosphatidylinositols