Identification of a Mutant DNA Polymerase in Saccharomyces cerevisiae With an Antimutator Phenotype for Frameshift Mutations

Corresponding author: Linda J. Reha-Krantz, Department of Biological Sciences, CW405 BioSciences Bldg., University of Alberta, Edmonton, Alberta T6G 2E9, Canada., lreha{at}gpu.srv.ualberta.ca (E-mail)

Communicating editor: S. SANDMEYER

We propose that a ß-turn-ß structure, which plays a critical role in exonucleolytic proofreading in the bacteriophage T4 DNA polymerase, is also present in the Saccharomyces cerevisiae DNA pol . Site-directed mutagenesis was used to test this proposal by introducing a mutation into the yeast POL3 gene in the region that encodes the putative ß-turn-ß structure. The mutant DNA pol has a serine substitution in place of glycine at position 447. DNA replication fidelity of the G447S-DNA pol was determined in vivo by using reversion and forward assays. An antimutator phenotype for frameshift mutations in short homopolymeric tracts was observed for the G447S-DNA pol in the absence of postreplication mismatch repair, which was produced by inactivation of the MSH2 gene. Because the G447S substitution reduced frameshift but not base substitution mutagenesis, some aspect of DNA polymerase proofreading appears to contribute to production of frameshifts. Possible roles of DNA polymerase proofreading in frameshift mutagenesis are discussed.

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