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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Biochemical basis for the essential genetic requirements of RecA and the β-clamp in Pol V activation
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Biochemical basis for the essential genetic requirements of RecA and the β-clamp in Pol V activation

机译：RecV和β-钳位基因在Pol V激活中基本遗传要求的生化基础

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摘要

In Escherichia colt, it is genetically well established that the β-clamp and RecA are essential cofactors that endow DNA polymerase (Pol)rnV with lesion bypass activity. However, the biochemical basis for these requirements is still largely unknown. Because the process of translesion synthesis (TLS) requires that the specialized ONA polymerase synthesize in a single binding event a TLS patch that is long enough to resist external proofreading, it is critical to monitor PolrnV burst synthesis. Here, we dissect the distinct roles that RecA and the β-clamp perform during the Pol V activation process using physiologically relevant long single-stranded template DNA, similar to those used in genetic assays. Our data show that the β-clamp endows the complex between Pol V and the template DNA with increased stability. Also, the RecA filament formed in cis on the single-stranded DNA produced downstream from the lesion stretches the template DNA to allow smooth elongation of the nascent strand by Pol V. The concurrent action of both cofactors is required for achieving productive TLS events. The present article presents an integrated view of TLS under physiologically relevant conditions in F. coli that may represent a paradigm for lesion bypass in other organisms.
机译：在大肠杆菌中，从基因上已经很好地证明了β钳和RecA是赋予DNA聚合酶（Pol）rnV具有损伤旁路活性的必要辅助因子。但是，这些要求的生化基础仍是未知之数。因为跨病变合成（TLS）的过程要求专门的ONA聚合酶在单个结合事件中合成足够长的TLS补丁以抵抗外部校对，所以监视PolrnV突发合成至关重要。在这里，我们使用生理学上相关的长单链模板DNA来解析RecA和β-钳位在Pol V激活过程中所发挥的独特作用，这与基因检测中所使用的相似。我们的数据表明，β-钳位赋予了Pol V和模板DNA之间的复合物更高的稳定性。同样，在病变下游产生的单链DNA上顺式形成的RecA细丝拉伸了模板DNA，从而允许Pol V平滑地延伸新生链。实现生产性TLS事件需要两个辅因子同时起作用。本文介绍了在大肠杆菌中生理相关条件下的TLS的完整视图，这可能代表了其他生物体中病变绕过的范例。

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    Shingo Fujii; Robert P. Fuchs;

  • 作者单位

    Centre National de la Recherche Scientifique, Unite Propre de Recherche 3081, Genome Instability and Carcinogenesis, Conventionne par I'Universite d'Aix-Marseille 2, 31, chemin Joseph Aiguier, 13402 Marseille cedex 20, France;

    Centre National de la Recherche Scientifique, Unite Propre de Recherche 3081, Genome Instability and Carcinogenesis, Conventionne par I'Universite d'Aix-Marseille 2, 31, chemin Joseph Aiguier, 13402 Marseille cedex 20, France;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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  • 正文语种 eng
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