Beyond translesion synthesis: polymerase κ fidelity as a potential determinant of microsatellite stability.
Beyond translesion synthesis: polymerase κ fidelity as a potential determinant of microsatellite stability.
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DOI：
10.1093/nar/gkr889
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发表时间：
2012-02
影响因子：
14.9
通讯作者：
Eckert KA
中科院分区：
文献类型：
--
作者：
Hile SE;Wang X;Lee MY;Eckert KA
微卫星DNA合成是人类基因组复制的重要组成部分，必须忠实地发生。然而，酵母复制型DNA聚合酶不具有高保真的微卫星合成。我们假设Y-家族聚合酶的结构特征，促进准确的translesion合成可能会促进准确的微卫星合成。我们使用体外HSV-tk测定比较了人聚合酶κ（Pol κ）和η（Pol η）与复制型人聚合酶δ全酶（Pol δ4）的结晶度。HSV-tk基因内部2-3个单位重复内插入/缺失（indel）错误的相对聚合酶准确度与文献一致：Pol δ4 >> Pol κ或Pol η。相比之下，[GT]10和[TC]11微卫星内基于单位的indel错误的相对聚合酶准确度为：Pol κ ≥ Pol δ4 > Pol η。使用[GT]模板时，Pos κ和δ4之间的差异幅度最大。在生物化学上，Pol κ在[GT]等位基因内的合成终止比Pol δ4少。在双重聚合酶反应中，Pol κ与停滞或移动的Pol δ4竞争，从而减少终止。我们的研究结果挑战了pol κ容易出错的思想，并表明具有互补生化特性的DNA聚合酶可以在重复序列上协同发挥作用。
Microsatellite DNA synthesis represents a significant component of human genome replication that must occur faithfully. However, yeast replicative DNA polymerases do not possess high fidelity for microsatellite synthesis. We hypothesized that the structural features of Y-family polymerases that facilitate accurate translesion synthesis may promote accurate microsatellite synthesis. We compared human polymerases κ (Pol κ) and η (Pol η) fidelities to that of replicative human polymerase δ holoenzyme (Pol δ4), using the in vitro HSV-tk assay. Relative polymerase accuracy for insertion/deletion (indel) errors within 2–3 unit repeats internal to the HSV-tk gene concurred with the literature: Pol δ4 >> Pol κ or Pol η. In contrast, relative polymerase accuracy for unit-based indel errors within [GT]10 and [TC]11 microsatellites was: Pol κ ≥ Pol δ4 > Pol η. The magnitude of difference was greatest between Pols κ and δ4 with the [GT] template. Biochemically, Pol κ displayed less synthesis termination within the [GT] allele than did Pol δ4. In dual polymerase reactions, Pol κ competed with either a stalled or moving Pol δ4, thereby reducing termination. Our results challenge the ideology that pol κ is error prone, and suggest that DNA polymerases with complementary biochemical properties can function cooperatively at repetitive sequences.
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DOI：
10.1073/pnas.0902175106
发表时间：
2009-06-16
影响因子：
11.1
作者：
Acharya, Narottam;Johnson, Robert E.;Prakash, Satya
通讯作者：
Prakash, Satya
影响因子：
64.8
作者：
Jarosz, DF;Godoy, VG;Walker, GC
通讯作者：
Walker, GC
影响因子：
14.9
作者：
Korona DA;Lecompte KG;Pursell ZF
通讯作者：
Pursell ZF
影响因子：
2.9
作者：
Eckert, KA;Mowery, A;Hile, SE
通讯作者：
Hile, SE
DOI：
10.1073/pnas.0901403106
发表时间：
2009-04-14
影响因子：
11.1
作者：
Indiani, Chiara;Langston, Lance D.;O'Donnell, Mike
通讯作者：
O'Donnell, Mike