Haracska Lajos, Prakash Satya, Prakash Louise
Sealy Center for Molecular Science, University of Texas Medical Branch, Galveston, Texas 77555-1061, USA.
Mol Cell Biol. 2003 Feb;23(4):1453-9. doi: 10.1128/MCB.23.4.1453-1459.2003.
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.
酿酒酵母中的遗传学研究表明,DNA聚合酶(Pol)ζ是紫外线及其他DNA损伤剂诱导诱变所必需的。然而,Pol ζ自身在通过DNA损伤进行复制时效率极低;相反,它通过从另一种DNA聚合酶插入损伤对面的核苷酸处延伸来促进损伤的诱变跨越。到目前为止,Pol ζ在环丁烷嘧啶二聚体、(6-4)二嘧啶光产物和无碱基位点上的这种作用已得到证实。在此,我们研究Pol ζ是否能通过7,8-二氢-8-氧代鸟嘌呤(8-氧代G)和O(6)-甲基鸟嘌呤(m6G)损伤进行复制。我们选择这两种损伤进行此项研究,是因为复制性聚合酶Pol δ能够通过它们进行复制,尽管效率很低。我们发现,Pol ζ在这两种损伤对面插入核苷酸的效率都非常低,但它能有效地从Pol δ插入损伤对面的核苷酸处延伸。此外,当Pol δ与Pol ζ结合时,8-氧代G和m6G损伤的跨越效率最高,这表明Pol ζ在从Pol δ插入损伤对面的核苷酸处延伸中发挥作用。因此,Pol ζ是一种高度特化的聚合酶,它能够从与DNA损伤相对的引物末端高效延伸,而不论损伤的几何扭曲程度如何。然而,Pol ζ对模板残基的几何扭曲异常敏感,因为它在掺入核苷酸时效率极低,即使是与中等扭曲的8-氧代G和m6G损伤相对时也是如此。