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首页> 外文期刊>Cell discovery. >ATPase activity tightly regulates RecA nucleofilaments to promote homologous recombination
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ATPase activity tightly regulates RecA nucleofilaments to promote homologous recombination

机译：ATPase活性紧密调节RecA核丝以促进同源重组

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Homologous recombination (HR), catalyzed in an evolutionarily conserved manner by active RecA/Rad51 nucleofilaments, maintains genomic integrity and promotes biological evolution and diversity. The structures of RecA/Rad51 nucleofilaments provide information critical for the entire HR process. By exploiting a unique capillary electrophoresis-laser-induced fluorescence polarization assay, we have discovered an active form of RecA nucleofilament, stimulated by ATP hydrolysis, that contains mainly unbound nucleotide sites. This finding was confirmed by a nuclease protection assay and electron microscopy (EM) imaging. We further found that these RecA-unsaturated filaments promote strand exchange in vitro and HR in vivo . RecA mutants (P67D and P67E), which only form RecA-unsaturated nucleofilaments, were able to mediate HR in vitro and in vivo , but mutants favoring the formation of the saturated nucleofilaments failed to support HR. We thus present a new model for RecA-mediated HR in which RecA utilizes its intrinsic DNA binding-dependent ATPase activity to remodel the nucleofilaments to a less saturated form and thereby promote HR.
机译：同源重组（HR），由活性RecA / Rad51核丝以进化保守的方式催化，可保持基因组完整性并促进生物学进化和多样性。 RecA / Rad51核丝的结构提供了对整个HR过程至关重要的信息。通过利用独特的毛细管电泳-激光诱导的荧光极化测定，我们发现了一种活性形式的RecA核丝，受ATP水解刺激，主要包含未结合的核苷酸位点。核酸酶保护分析和电子显微镜（EM）成像证实了这一发现。我们进一步发现，这些RecA不饱和细丝促进体外链交换和体内HR。仅形成RecA不饱和核丝的RecA突变体（P67D和P67E）能够在体内和体外介导HR，但是偏爱饱和核丝形成的突变体却无法支持HR。因此，我们提出了RecA介导的HR的新模型，其中RecA利用其内在的DNA结合依赖性ATPase活性将核丝重塑成饱和度较低的形式，从而促进HR。

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