Genetics, Vol 122, 535-542, Copyright © 1989

INVESTIGATIONS

Disruption of the RAD52 Gene Alters the Spectrum of Spontaneous SUP4-o Mutations in Saccharomyces cerevisiae

B. A. Kunz, M. G. Peters, S. E. Kohalmi, J. D. Armstrong, M. Glattke and K. Badiani
Microbiology Department, The University of Manitoba, Winnepeg, Manitoba, Canada R3T 2N2

Defects in the RAD52 gene of the yeast Saccharomyces cerevisiae confer a mutator phenotype. To characterize this effect in detail, a collection of 238 spontaneous SUP4-o mutations arising in a strain having a disrupted RAD52 gene was analyzed by DNA sequencing. The resulting mutational spectrum was compared to that derived from an examination of 222 spontaneous mutations selected in a near-isogenic wild-type (RAD52) strain. This comparison revealed that the mutator phenotype was associated with an increase in the frequency of base-pair substitutions. All possible types of substitution were detected but there was a reduction in the relative fraction of A{middot}T -> G{middot}C transitions and an increase in the proportion of G{middot}C -> C{middot}G transversions. These changes were sufficient to cause a twofold greater preference for substitutions at G{middot}C sites in the rad52 strain despite a decrease in the fraction of G{middot}C -> T{middot}A transversions. There were also considerable differences between the distributions of substitutions within the SUP4-o gene. Base-pair changes occurred at fewer sites in the rad52 strain but the mutated sites included several that were not detected in the RAD52 background. Only two of the four sites that were mutated most frequently in the rad52 strain were also prominent in the wild-type strain and mutation frequencies at almost all sites common to both strains were greater for the rad52 derivative. Although single base-pair deletions occurred in the two strains with similar frequencies, several classes of mutation that were recovered in the wild-type background including multiple base-pair deletions, insertions of the yeast transposable element Ty, and more complex changes, were not detected in the rad52 strain. These results are discussed in relation to possible mechanisms that might account for the rad52 mutator effect.

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