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Recombinational repair of radiation-induced double-strand breaks occurs in the absence of extensive resection

机译:在没有大范围切除的情况下进行放射诱导的双链断裂的重组修复

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摘要

Recombinational repair provides accurate chromosomal restitution after double-strand break (DSB) induction. While all DSB recombination repair models include 5′-3′ resection, there are no studies that directly assess the resection needed for repair between sister chromatids in G-2 arrested cells of random, radiation-induced ‘dirty’ DSBs. Using our Pulse Field Gel Electrophoresis-shift approach, we determined resection at IR-DSBs in WT and mutants lacking exonuclease1 or Sgs1 helicase. Lack of either reduced resection length by half, without decreased DSB repair or survival. In the exo1Δ sgs1Δ double mutant, resection was barely detectable, yet it only took an additional hour to achieve a level of repair comparable to WT and there was only a 2-fold dose-modifying effect on survival. Results with a Dnl4 deletion strain showed that remaining repair was not due to endjoining. Thus, similar to what has been shown for a single, clean HO-induced DSB, a severe reduction in resection tract length has only a modest effect on repair of multiple, dirty DSBs in G2-arrested cells. Significantly, this study provides the first opportunity to directly relate resection length at DSBs to the capability for global recombination repair between sister chromatids.
机译:重组修复可在诱导双链断裂(DSB)后提供准确的染色体修复。尽管所有DSB重组修复模型均包括5'-3'切除,但尚无直接评估在辐射诱发的“脏” DSB的G-2停滞细胞中姊妹染色单体之间进行修复所需的切除的研究。使用我们的脉冲场凝胶电泳移位方法,我们确定了WT和缺少外切核酸酶1或Sgs1解旋酶的突变体在IR-DSB处的切除。既不能将切除长度减少一半,也不能减少DSB的修复或存活率。在exo1Δsgs1Δ双突变体中,几乎无法检测到切除,但仅需额外的一个小时即可达到与WT相当的修复水平,并且对存活率只有2倍的剂量调节作用。 Dnl4缺失菌株的结果表明,剩余的修复不是由于末端连接。因此,类似于单个清洁的HO诱导的DSB所显示的结果,切除道长度的严重减少仅对修复G2细胞中的多个脏DSB具有适度的作用。重要的是,这项研究提供了第一个机会,可以直接将DSB的切除长度与姐妹染色单体之间的整体重组修复能力联系起来。

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