Gibbs Peter E M, McDonald John, Woodgate Roger, Lawrence Christopher W
Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA.
Genetics. 2005 Feb;169(2):575-82. doi: 10.1534/genetics.104.034611. Epub 2004 Nov 1.
We have investigated the relative roles in vivo of Saccharomyces cerevisiae DNA polymerase eta, DNA polymerase zeta, Rev1 protein, and the DNA polymerase delta subunit, Pol32, in the bypass of an abasic site, T-T (6-4) photoadduct and T-T cis-syn cyclobutane dimer, by transforming strains deleted for RAD30, REV3, REV1, or POL32 with duplex plasmids carrying one of these DNA lesions located within a 28-nucleotide single-stranded region. DNA polymerase eta was found to be involved only rarely in the bypass of the T-T (6-4) photoadduct or the abasic sites in the sequence context used, although, as expected, it was solely responsible for the bypass of the T-T dimer. We argue that DNA polymerase zeta, rather than DNA polymerase delta as previously suggested, is responsible for insertion in bypass events other than those in which polymerase eta performs this function. However, DNA polymerase delta is involved indirectly in mutagenesis, since the strain lacking its Pol32 subunit, known to be deficient in mutagenesis, shows as little bypass of the T-T (6-4) photoadduct or the abasic sites as those deficient in Pol zeta or Rev1. In contrast, bypass of the T-T dimer in the pol32delta strain occurs at the wild-type frequency.
我们通过用携带位于28个核苷酸单链区域内的这些DNA损伤之一的双链质粒转化缺失RAD30、REV3、REV1或POL32的菌株,研究了酿酒酵母DNA聚合酶η、DNA聚合酶ζ、Rev1蛋白和DNA聚合酶δ亚基Pol32在体内绕过无碱基位点、T-T(6-4)光产物和T-T顺式-顺环丁烷二聚体中的相对作用。发现DNA聚合酶η在所用序列背景下很少参与绕过T-T(6-4)光产物或无碱基位点,尽管正如预期的那样,它完全负责绕过T-T二聚体。我们认为,负责在聚合酶η执行此功能以外的绕过事件中进行插入的是DNA聚合酶ζ,而不是先前认为的DNA聚合酶δ。然而,DNA聚合酶δ间接参与诱变,因为已知缺乏其Pol32亚基且诱变缺陷的菌株对T-T(6-4)光产物或无碱基位点的绕过与缺乏Pol ζ或Rev1的菌株一样少。相比之下,pol32δ菌株中T-T二聚体的绕过以野生型频率发生。