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首页> 外文期刊>Molecular and Cellular Biology >Palm Mutants in DNA Polymerases α and η Alter DNA Replication Fidelity and Translesion Activity
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Palm Mutants in DNA Polymerases α and η Alter DNA Replication Fidelity and Translesion Activity

机译：DNA聚合酶α和η中的棕榈突变体改变DNA复制的保真度和转移活性

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We isolated active mutants in Saccharomyces cerevisiae DNA polymerase α that were associated with a defect in error discrimination. Among them, L868F DNA polymerase α has a spontaneous error frequency of 3 in 100 nucleotides and 570-fold lower replication fidelity than wild-type (WT) polymerase α. In vivo, mutant DNA polymerases confer a mutator phenotype and are synergistic with msh2 or msh6, suggesting that DNA polymerase α-dependent replication errors are recognized and repaired by mismatch repair. In vitro, L868F DNA polymerase α catalyzes efficient bypass of a cis-syn cyclobutane pyrimidine dimer, extending the 3′ T 26,000-fold more efficiently than the WT. Phe34 is equivalent to residue Leu868 in translesion DNA polymerase η, and the F34L mutant of S. cerevisiae DNA polymerase η has reduced translesion DNA synthesis activity in vitro. These data suggest that high-fidelity DNA synthesis by DNA polymerase α is required for genomic stability in yeast. The data also suggest that the phenylalanine and leucine residues in translesion and replicative DNA polymerases, respectively, might have played a role in the functional evolution of these enzyme classes.
机译：我们在啤酒酵母DNA聚合酶α中分离了与错误鉴别缺陷有关的活性突变体。其中，L868F DNA聚合酶α的自发错误频率为100个核苷酸中的3个，并且复制保真度比野生型（WT）聚合酶α低570倍。在体内，突变的DNA聚合酶具有突变表型，并且与 msh2 或 msh6 协同作用，表明DNA聚合酶α依赖性复制错误可以通过错配修复得到识别和修复。在体外，L868F DNA聚合酶α催化顺式-syn 环丁烷嘧啶二聚体的有效旁路，其3'T的延伸效率比WT高26,000倍。 Phe34等同于跨病变DNA聚合酶η中的残基Leu868和 S的F34L突变体。酿酒酵母DNA聚合酶η在体外降低了病灶的DNA合成活性。这些数据表明，DNA聚合酶α进行高保真DNA合成是酵母基因组稳定性所必需的。数据还表明，跨病变和复制型DNA聚合酶中的苯丙氨酸和亮氨酸残基可能分别在这些酶类别的功能进化中起作用。

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