Mutations in Cognate Genes of Saccharomyces cerevisiae hsp70 Result in Reduced Growth Rates at Low Temperatures

Abstract

Expression of two Saccharomyces cerevisiae genes (YG101 and YG103) that are related to the gene encoding inducible 70K protein (hsp70) is repressed upon heat shock. Mutations of the two genes were constructed in vitro and substituted into the yeast genome in place of the wild-type alleles. No phenotypic effect of single mutations of either gene was detected. However, cells containing both YG101 and YG103 mutations showed altered growth properties; double-mutation cells possess an optimal growth temperature of 37°C rather than 30°C and grow increasingly poorly as the temperature is lowered. Mutations of two other members of this hsp70-related multigene family, YG100 and YG102, have been analyzed (E. A. Craig and K. Jacobsen, Cell 38:841–849, 1984). Cells containing both YG100 and YG102 mutations cannot form colonies at 37°C. Fusions between the YG101 and YG102 promoter regions and the YG100 and YG101 structural genes, respectively, were constructed. The YG101 promoter-YG100 structural gene fusion was not able to restore normal growth properties to the yg101⁻ yg103⁻ mutant. Also, yg100⁻ yg102⁻ cells containing the YG102 promoter-YG101 structural gene fusion were unable to grow at 37°C. Failure of the protein products of related genes to rescue the relative cold sensitivity of growth suggests that members of the hsp70 multigene family are functionally distinct.