Isolation of a Fission Yeast Mutant Cell Affected in MAP Kinase Signaling and Sterol Biosynthesis

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We have previously demonstrated that calcineurin and the Pmk1 MAP kinasepathway play an antagonistic role in Cl– homeostasis. Using this relationship, wescreened for mutations that show vic (viable in the presence of immunosuppressant andchloride ion) phenotype and isolated a vic6 mutant cell.The vic6 mutant cells also showed sensitivity to high temperature. Using thisphenotype, we isolated hmg1+ gene, encoding a HMG-CoA reductase. Consistently, thevic6 mutant cells exhibited hypersensitivity to miconazole, an inhibitor of ergosterolbiosynthesis and showed aberrant intracellular localization of filipin, suggesting thatthe mutant cells are affected in the sterol biosynthesis. In addition, overexpression ofthe hmg1+ gene complemented the phenotype of vic1-1/cpp1-v1 mutant cells, an allele ofthe gene encoding a farnesyltransferase, whereas overexpression of the cpp1+ geneexacerbated the temperature-sensitive phenotype of the vic6 mutant cells.