Veaute Xavier, Jeusset Josette, Soustelle Christine, Kowalczykowski Stephen C, Le Cam Eric, Fabre Francis
CEA, DSV, Département de Radiobiologie et Radiopathologie, UMR217 CNRS/CEA, BP6, 92265 Fontenay aux Roses Cedex, France.
Nature. 2003 May 15;423(6937):309-12. doi: 10.1038/nature01585.
Homologous recombination is a ubiquitous process with key functions in meiotic and vegetative cells for the repair of DNA breaks. It is initiated by the formation of single-stranded DNA on which recombination proteins bind to form a nucleoprotein filament that is active in searching for homology, in the formation of joint molecules and in the exchange of DNA strands. This process contributes to genome stability but it is also potentially dangerous to cells if intermediates are formed that cannot be processed normally and thus are toxic or generate genomic rearrangements. Cells must therefore have developed strategies to survey recombination and to prevent the occurrence of such deleterious events. In Saccharomyces cerevisiae, genetic data have shown that the Srs2 helicase negatively modulates recombination, and later experiments suggested that it reverses intermediate recombination structures. Here we show that DNA strand exchange mediated in vitro by Rad51 is inhibited by Srs2, and that Srs2 disrupts Rad51 filaments formed on single-stranded DNA. These data provide an explanation for the anti-recombinogenic role of Srs2 in vivo and highlight a previously unknown mechanism for recombination control.
同源重组是一个普遍存在的过程,在减数分裂细胞和营养细胞中对于DNA断裂的修复具有关键作用。它由单链DNA的形成引发,重组蛋白结合在单链DNA上形成核蛋白丝,该核蛋白丝在寻找同源性、形成联合分子以及DNA链交换过程中发挥作用。这一过程有助于基因组稳定性,但如果形成的中间体不能正常处理,从而具有毒性或导致基因组重排,那么对细胞来说也是潜在危险的。因此,细胞必须已经发展出监测重组并防止此类有害事件发生的策略。在酿酒酵母中,遗传数据表明Srs2解旋酶对重组具有负调控作用,后来的实验表明它能逆转中间重组结构。在这里我们表明,Srs2抑制了由Rad51在体外介导的DNA链交换,并且Srs2破坏了在单链DNA上形成的Rad51丝。这些数据为Srs2在体内的抗重组作用提供了解释,并突出了一种以前未知的重组控制机制。