Yeast DNA polymerase zeta 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 zeta 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 zeta has been established for cyclobutane pyrimidine dimers, (6-4) dipyrimidine photoproducts, and abasic sites. Here, we examine whether Pol zeta 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, Pol delta, can replicate through them, albeit weakly. We found that Pol zeta is very inefficient at inserting nucleotides opposite both these lesions, but it can efficiently extend from the nucleotides inserted opposite them by Pol delta. Also, the most efficient bypass of 8-oxoG and m6G lesions occurs when Pol delta is combined with Pol zeta, indicating a role for Polzeta in extending from the nucleotides inserted opposite these lesions by Pol delta. Thus, Pol zeta is a highly specialized polymerase that can proficiently extend from the primer ends opposite DNA lesions, irrespective of their degree of geometric distortion. Pol zeta, 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.
Haracska
Lajos
L
Sealy Center for Molecular Science, University of Texas Medical Branch, Galveston, Texas 77555-1061, USA.
Prakash
Satya
S
Prakash
Louise
L
D001483:Base Sequence; D017931:DNA Primers; D004260:DNA Repair / Q000502:physiology; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D006147:Guanine / Q000031:analogs & derivatives* / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity
C453560:7,8-dihydro-8-oxoguanine; D017931:DNA Primers; D029701:Saccharomyces cerevisiae Proteins; D006147:Guanine; C008449:O-(6)-methylguanine; C425929:DNA polymerase zeta; D004259:DNA-Directed DNA Polymerase