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首页> 外文期刊>Nature >RecA bundles mediate homology pairing between distant sisters during DNA break repair
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RecA bundles mediate homology pairing between distant sisters during DNA break repair

机译：RecA束在DNA断裂修复过程中介导遥远姐妹之间的同源配对

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DNA double-strand break (DSB) repair by homologous recombination has evolved to maintain genetic integrity in all organisms1. Although many reactions that occur during homologous recombination are known, it is unclear where, when and how they occur in cells. Here, by using conventional and super-resolution microscopy, we describe the progression of DSB repair in live Escherichia coli. Specifically, we investigate whether homologous recombination can occur efficiently between distant sister loci that have segregated to opposite halves of an E. coli cell. We show that a site-specific DSB in one sister can be repaired efficiently using distant sister homology. After RecBCD processing of the DSB, RecA is recruited to the cut locus, where it nucleates into a bundle that contains many more RecA molecules than can associate with the two single-stranded DNA regions that form at the DSB. Mature bundles extend along the long axis of the cell, in the space between the bulk nucleoid and the inner membrane. Bundle formation is followed by pairing, in which the two ends of the cut locus relocate at the periphery of the nucleoid and together move rapidly towards the homology of the uncut sister. After sister locus pairing, RecA bundles disassemble and proteins that act late in homologous recombination are recruited to give viable recombinants 1 -2-generation-time equivalents after formation of the initial DSB. Mutated RecA proteins that do not form bundles are defective in sister pairing and in DSB-induced repair. This work reveals an unanticipated role of RecA bundles in channelling the movement of the DNA DSB ends, thereby facilitating the long-range homology search that occurs before the strand invasion and transfer reactions.
机译：通过同源重组修复DNA双链断裂（DSB）的方法已经发展到可以维持所有生物的遗传完整性。尽管已知在同源重组期间发生的许多反应，但不清楚它们在细胞中的何处，何时以及如何发生。在这里，通过使用常规和超高分辨率显微镜，我们描述了在活大肠杆菌中DSB修复的进展。具体来说，我们调查是否已经可以分离到大肠杆菌细胞的相反两半的远距离姐妹基因座之间有效地发生同源重组。我们表明，可以使用远距离姐妹同源性有效修复一个姐妹中特定于位点的DSB。在对DSB进行RecBCD处理后，RecA被募集到切割位点，在那里成核成束，该束包含的RecA分子比与DSB处形成的两个单链DNA区域关联的更多。成熟的束在本体核与内膜之间的空间中沿细胞的长轴延伸。束形成之后是配对，其中切割的基因座的两个末端位于核苷酸的外围，并一起迅速向未切割的姐妹的同源性移动。在姐妹基因座配对后，RecA束分解，并在同源重组中起后期作用的蛋白质被募集，以在形成初始DSB后获得具有活力的1-2代时间的重组体。不形成束的突变的RecA蛋白在姐妹配对和DSB诱导的修复中有缺陷。这项工作揭示了RecA束在引导DNA DSB末端运动中的出乎意料的作用，从而促进了在链入侵和转移反应之前发生的远程同源性搜索。

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  • 来源
    《Nature》 |2014年第7487期|249-253|共5页
  • 作者

    Christian Lesterlin; Graeme Ball; Lothar Schermelleh; David J. Sherratt;

  • 作者单位

    Department of Biochemistry, University of Oxford, Oxford 0X1 3QU, UK;

    Department of Biochemistry, University of Oxford, Oxford 0X1 3QU, UK;

    Department of Biochemistry, University of Oxford, Oxford 0X1 3QU, UK;

    Department of Biochemistry, University of Oxford, Oxford 0X1 3QU, UK;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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