Yeast DNA polymerase ξ is an efficient extender of primer ends opposite from 7,8-dihydro-8-oxoguanine and O6-methylguanine

Genetic studies in Saccharomyces cerevisiae have indicated the requirement of DNA polymerase (Pol) zeta for mutagenesis induced by UV light and by other DNA damaging agents. However, on its own, Pol is highly inefficient at replicating through DNA lesions; rather, it promotes their mutagenic bypass by extending from the nucleotide inserted opposite the lesion by another DNA polymerase. So far, such a role for Pol has been established for cyclobutane pyrimidine dimers, (6-4) dipyrimidine photoproducts, and abasic sites. Here, we examine whether Pol can replicate through the 7,8-dihydro-8-oxoguanine (8-oxoG) and O-6-methylguanine (m6G) lesions. We chose these two lesions for this study because the replicative polymerase, Poldelta, can replicate through them, albeit weakly. We found that Pol is very inefficient at inserting nucleotides opposite both these lesions, but it can efficiently extend from the nucleotides inserted opposite them by Poldelta. Also, the most efficient bypass of 8-oxoG and m6G lesions occurs when Poldelta is combined with Pol, indicating a role for Polzeta in extending from the nucleotides inserted opposite these lesions by Polzeta. Thus, Pozeta is a highly specialized polymerase that can proficiently extend from the primer ends opposite DNA lesions, irrespective of their degree of geometric distortion. Polzeta, however, is unusually sensitive to geometric distortion of the templating residue, as it is highly inefficient at incorporating nucleotides even opposite the moderately distorting 8-oxoG and m6G lesions.