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首页> 美国卫生研究院文献>Molecular and Cellular Biology >DEAD Box 1 Facilitates Removal of RNA and Homologous Recombination at DNA Double-Strand Breaks
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DEAD Box 1 Facilitates Removal of RNA and Homologous Recombination at DNA Double-Strand Breaks

机译：DEAD Box 1有助于在DNA双链断裂时去除RNA和同源重组。

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摘要

Although RNA and RNA-binding proteins have been linked to double-strand breaks (DSBs), little is known regarding their roles in the cellular response to DSBs and, if any, in the repair process. Here, we provide direct evidence for the presence of RNA-DNA hybrids at DSBs and suggest that binding of RNA to DNA at DSBs may impact repair efficiency. Our data indicate that the RNA-unwinding protein DEAD box 1 (DDX1) is required for efficient DSB repair and cell survival after ionizing radiation (IR), with depletion of DDX1 resulting in reduced DSB repair by homologous recombination (HR). While DDX1 is not essential for end resection, a key step in homology-directed DSB repair, DDX1 is required for maintenance of the single-stranded DNA once generated by end resection. We show that transcription deregulation has a significant effect on DSB repair by HR in DDX1-depleted cells and that RNA-DNA duplexes are elevated at DSBs in DDX1-depleted cells. Based on our combined data, we propose a role for DDX1 in resolving RNA-DNA structures that accumulate at DSBs located at sites of active transcription. Our findings point to a previously uncharacterized requirement for clearing RNA at DSBs for efficient repair by HR.
机译：尽管RNA和RNA结合蛋白已经与双链断裂（DSB）相连，但人们对其在DSB的细胞反应以及修复过程中的作用所知甚少。在这里，我们提供了DSB处RNA-DNA杂合体存在的直接证据，并建议在DSB处RNA与DNA的结合可能会影响修复效率。我们的数据表明，解离RNA的蛋白质DEAD盒1（DDX1）是有效的DSB修复和电离辐射（IR）后细胞存活所必需的，DDX1的耗尽导致通过同源重组（HR）导致DSB修复减少。尽管DDX1对于末端切除不是必不可少的，这是同源性指导的DSB修复的关键步骤，但一旦末端切除产生了单链DNA，则需要DDX1来维持单链DNA。我们显示，转录失调对DDX1缺失的细胞中HR的DSB修复有重要影响，并且RNA-DNA双链体在DDX1缺失的细胞中的DSBs处升高。根据我们的综合数据，我们提出了DDX1在解析RNA-DNA结构中的作用，该结构积累在位于主动转录位点的DSB处。我们的研究结果指出，以前对于清除DSB上的RNA以进行HR有效修复的要求不高。

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