Department of Chemistry and Biochemistry, New Mexico State University, P.O. Box 30001, MSC 3C, Las Cruces, New Mexico 88003, USA.
Nat Commun. 2017 May 17;8:15282. doi: 10.1038/ncomms15282.
RecN is a cohesin-like protein involved in DNA double-strand break repair in bacteria. The RecA recombinase functions to mediate repair via homologous DNA strand invasion to form D-loops. Here we provide evidence that the RecN protein stimulates the DNA strand invasion step of RecA-mediated recombinational DNA repair. The intermolecular DNA tethering activity of RecN protein described previously cannot fully explain this novel activity since stimulation of RecA function is species-specific and requires RecN ATP hydrolysis. Further, DNA-bound RecA protein increases the rate of ATP hydrolysis catalysed by RecN during the DNA pairing reaction. DNA-dependent RecN ATPase kinetics are affected by RecA protein in a manner suggesting a specific order of protein-DNA assembly, with RecN acting after RecA binds DNA. We present a model for RecN function that includes presynaptic stimulation of the bacterial repair pathway perhaps by contributing to the RecA homology search before ternary complex formation.
RecN 是一种类似黏合蛋白的蛋白质,参与细菌中的 DNA 双链断裂修复。RecA 重组酶通过同源 DNA 链入侵来形成 D 环,从而介导修复。在这里,我们提供的证据表明 RecN 蛋白可刺激 RecA 介导的重组 DNA 修复的 DNA 链入侵步骤。先前描述的 RecN 蛋白的分子间 DNA 系链活性不能完全解释这种新的活性,因为 RecA 功能的刺激是具有物种特异性的,并且需要 RecN 的 ATP 水解。此外,结合 DNA 的 RecA 蛋白在 DNA 配对反应中增加了 RecN 催化的 ATP 水解的速率。DNA 依赖性 RecN ATPase 动力学受 RecA 蛋白的影响,表明蛋白-DNA 组装的特定顺序,RecA 结合 DNA 后 RecN 起作用。我们提出了 RecN 功能的模型,该模型包括在三元复合物形成之前,通过有助于 RecA 同源搜索来对细菌修复途径进行突触前刺激。