Competition between replicative and translesion polymerases during homologous recombination repair in Drosophila.
Competition between replicative and translesion polymerases during homologous recombination repair in Drosophila.
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DOI：
10.1371/journal.pgen.1002659
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发表时间：
2012
期刊：
影响因子：
4.5
通讯作者：
McVey M
中科院分区：
文献类型：
--
作者：
Kane DP;Shusterman M;Rong Y;McVey M
在后生动物中，DNA在同源重组修复双链断裂过程中合成的机制知之甚少。具体而言，进行修复合成的聚合酶的身份以及它们如何被募集到修复位点尚不清楚。在这里，我们研究了几种不同的聚合酶在果蝇同源重组修复过程中的作用。使用缺口修复试验，我们发现，同源重组受损的果蝇缺乏DNA聚合酶zeta，在较小程度上，聚合酶eta。此外，Pol32蛋白，聚合酶δ复合物的一部分，是需要广泛合成的修复所必需的。Rev1与多种跨损伤聚合酶相互作用，导致间隙修复期间合成增加。总之，我们的研究结果支持了一个模型，其中translesion聚合酶和聚合酶δ复合体在同源重组修复过程中竞争。此外，他们将Rev1确定为调节修复合成程度的关键因子。DNA聚合酶在DNA复制和各种类型的DNA修复过程中都是必需的。DNA双链断裂通常通过同源重组修复，同源重组是一种保守的过程，其中DNA从相似的模板复制到断裂位点。在DNA双链断裂的同源重组修复过程中起作用的特异性聚合酶在多细胞生物中还没有完全表征。在这项研究中，我们创建了缺乏一种或多种DNA聚合酶的果蝇突变株，并确定了它们在同源重组过程中合成大量DNA的能力。我们发现，易错translesion聚合酶eta和zeta在合成的起始过程中发挥重叠的作用，而Pol32亚基的复制聚合酶δ复合物是需要修复涉及大量的合成。此外，我们发现，缺乏Rev1 translesion聚合酶的果蝇在缺口修复过程中比正常果蝇合成更多的DNA。我们的研究结果表明，复制和translesion聚合酶都参与同源重组，并确定Rev1作为一种蛋白质，可以调节各种聚合酶的双链断裂修复中间体的访问。
In metazoans, the mechanism by which DNA is synthesized during homologous recombination repair of double-strand breaks is poorly understood. Specifically, the identities of the polymerase(s) that carry out repair synthesis and how they are recruited to repair sites are unclear. Here, we have investigated the roles of several different polymerases during homologous recombination repair in Drosophila melanogaster. Using a gap repair assay, we found that homologous recombination is impaired in Drosophila lacking DNA polymerase zeta and, to a lesser extent, polymerase eta. In addition, the Pol32 protein, part of the polymerase delta complex, is needed for repair requiring extensive synthesis. Loss of Rev1, which interacts with multiple translesion polymerases, results in increased synthesis during gap repair. Together, our findings support a model in which translesion polymerases and the polymerase delta complex compete during homologous recombination repair. In addition, they establish Rev1 as a crucial factor that regulates the extent of repair synthesis. DNA polymerases are required during both DNA replication and various types of DNA repair. DNA double-strand breaks are frequently repaired by homologous recombination, a conservative process in which DNA is copied into the break site from a similar template. The specific polymerases that operate during homologous recombination repair of DNA double-strand breaks have not been fully characterized in multicellular organisms. In this study, we created mutant strains of Drosophila lacking one or more DNA polymerases and determined their ability to synthesize large amounts of DNA during homologous recombination. We found that the error-prone translesion polymerases eta and zeta play overlapping roles during the initiation of synthesis, while the Pol32 subunit of the replicative polymerase delta complex is required for repair involving large amounts of synthesis. In addition, we showed that flies lacking the Rev1 translesion polymerase synthesize more DNA during gap repair than their normal counterparts. Our results demonstrate that both replicative and translesion polymerases are involved in homologous recombination and identify Rev1 as a protein that may regulate the access of various polymerases to double-strand break repair intermediates.
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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：
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Lehmann, Alan R.;Niimi, Atsuko;Green, Catherine M.
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