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A mechanistic basis for Mre11-directed DNA joining at microhomologies

机译:微量同源的Mre11定向DNA连接的机制基础

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摘要

Repair of DNA double-strand breaks in vertebrate cells occurs mainly by an end-joining process that often generates junctions with sequence homologies of a few nucleotides. Mre11 is critical for this mode of repair in budding yeast and has been implicated in the microhomology-based joining. Here, we show that Mre11 exonuclease activity is sensitive to the presence of heterologous DNA, and to the structure and sequence of its ends. Addition of mismatched DNA ends stimulates degradation of DNA by Mre11, whereas cohesive ends strongly inhibit it. Furthermore, if a sequence identity is revealed during the course of degradation, it causes Mre11 nuclease activity to pause, thus stabilizing the junction at a site of microhomology. A nuclease-deficient Mre11 mutant that still binds DNA can also stimulate degradation by wild-type Mre11, suggesting that Mre11-DNA complexes may interact to bridge DNA ends and facilitate DNA joining.
机译:脊椎动物细胞中DNA双链断裂的修复主要通过末端连接过程进行,该过程通常会产生与几个核苷酸的序列同源性的连接。 Mre11对于这种在发芽酵母中的修复模式至关重要,并且与基于微同源性的连接有关。在这里,我们显示Mre11核酸外切酶活性对异源DNA的存在及其末端的结构和序列敏感。添加错配的DNA末端会刺激Mre​​11降解DNA,而具有粘性的末端会强烈抑制它。此外,如果在降解过程中揭示了序列同一性,则它将导致Mre11核酸酶活性暂停,从而稳定微同源位点的连接。仍然结合DNA的核酸酶缺陷型Mre11突变体也可以刺激野生型Mre11降解,这表明Mre11-DNA复合物可能相互作用以桥接DNA末端并促进DNA连接。

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