Isolation and characterization of the aureobasidin A-resistant gene, aur1(R), on Schizosaccharomyces pombe: Roles of Aur1p+ in cell morphogenesis

Abstract

To study the mechanism of action of the antibiotic aureobasidin A (AbA) on yeasts, we isolated a dominant mutant of Schizosaccharomyces pombe which gave high resistance to AbA. From a genomic library of the mutant, an aur1(R) mutant gene conferring AbA resistance was isolated. One amino-acid mutation, a substitution of glycine with cysteine at residue 240, was responsible for the acquisition of AbA resistance. The wild-type aur1+ gene was essential for viability, and its over-expression enhanced significant resistance to AbA. The predicted protein of S. pombe aur1(R) was highly homologous in primary structure and hydropathy profile with that of Saccharomyces cerevisiae AUR1(R) isolated as an AbA-resistance gene. To analyze a role in cell growth of S. pombe aur1+, temperature-sensitive mutants (aur1(ts)) were obtained by random mutagenesis procedures using a modified PCR. The aur1(ts) mutation caused a defect in cell elongation at the non-permissive temperature and finally led to cell death. These results suggest that Aur1p was a target of the antibiotic AbA and was required in the cell elongation of cell-end tips and in the viability of S. pombe.