Original Articles

Molecular and General Genetics MGG

, Volume 242, Issue 5, pp 517-527

First online:

Temperature-sensitive mutants of hsp82 of the budding yeast Saccharomyces cerevisiae

  • Yoko KimuraAffiliated withDepartment of Cell Biology, Tokyo Metropolitan Institute of Medical ScienceInstitute for Molecular and Cellular Biology, Osaka University
  • , Seiji MatsumotoAffiliated withDepartment of Cell Biology, Tokyo Metropolitan Institute of Medical Science
  • , Ichiro YaharaAffiliated withDepartment of Cell Biology, Tokyo Metropolitan Institute of Medical Science

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Abstract

The budding yeast Saccharomyces cerevisiae has two HSP90-related genes per haploid genome, HSP82 and HSC82. Random mutations were induced in vitro in the HSP82 gene by treatment of the plasmid with hydroxylamine. Four temperature-sensitive (ts) mutants and one simultaneously is and cold-sensitivie (cs) mutant were then selected in a yeast strain in which HSC82 had previously been disrupted. The mutants were found to have single base changes in the coding region, which caused single amino acid substitutions in the HSP82 protein. All of these mutations occurred in amino acid residues that are well conserved among HSP90-related proteins of various species from Escherichia coli to human. Various properties including cell morphology, macromolecular syntheses and thermosensitivity were examined in each mutant at both the permissive and nonpermissive temperatures. The mutations in HSP82 caused pleiotropic effects on these properties although the phenotypes exhibited at the nonpermissive temperature varied among the mutants.

Key words

Saccharomyces cerevisiae HSP82 Random in vitro mutagenesis Temperature-sensitive mutants