Roles of RAD6 epistasis group members in spontaneous Polξ-dependent translesion synthesis in Saccharomyces cerevisiae

DNA lesions that arise during normal cellular metabolism can block the progress of replicative DNA polymerases, leading to cell cycle arrest and, in higher eukaryotes, apoptosis. Alternatively, Such blocking lesions can be temporarily tolerated rising either a recombination- or a translesion synthesis-based bypass mechanism. In Saccharomyces cerevisiae, members of the RAD6epistasis group are key players in the regulation of lesion bypass by the translesion DNA polymerase Pol zeta. In this study, changes in the reversion rate and spectrum of the lys2 Delta A746 -1 frameshift allele have been used to evaluate how the loss of members of the RAD6 epistasis group affects Pol zeta-dependent mutagenesis in response to spontaneous damage. Our data are consistent with a model in which Pol zeta-dependent mutagenesis relies on the presence of either Rad5 or Rad18, which promote two distinct error-prone pathways that partially overlap with respect to lesion specificity. The smallest subunit of Pol delta, Pol32, is also required for Pol zeta-dependent spontaneous mutagenesis, suggesting a cooperative role between Pol delta and Pol zeta for the bypass of spontaneous lesions. A third error-free pathway relies on the presence of Mms2, but may not require PCNA.