Muyrers J P, Zhang Y, Buchholz F, Stewart A F
Gene Expression Program, European Molecular Biology Laboratory, D-69117 Heidelberg, Germany.
Genes Dev. 2000 Aug 1;14(15):1971-82.
The initial steps of double-stranded break (DSB) repair by homologous recombination mediated by the 5'-3' exonuclease/annealing protein pairs, RecE/RecT and Redalpha/Redbeta, were analyzed. Recombination was RecA-independent and required the expression of both components of an orthologous pair, even when the need for exonuclease activity was removed by use of preresected substrates. The required orthologous function correlated with a specific protein-protein interaction, and recombination was favored by overexpression of the annealing protein with respect to the exonuclease. The need for both components of an orthologous pair was observed regardless of whether recombination proceeded via a single-strand annealing or a putative strand invasion mechanism. The DSB repair reactions studied here are reminiscent of the RecBCD/RecA reaction and suggest a general mechanism that is likely to be relevant to other systems, including RAD52 mediated recombination.
分析了由5'-3'核酸外切酶/退火蛋白对RecE/RecT和Redalpha/Redbeta介导的同源重组修复双链断裂(DSB)的初始步骤。重组不依赖RecA,并且需要直系同源对的两个组分都表达,即使通过使用预切除底物消除了对核酸外切酶活性的需求。所需的直系同源功能与特定的蛋白质-蛋白质相互作用相关,并且相对于核酸外切酶,退火蛋白的过表达有利于重组。无论重组是通过单链退火还是推测的链侵入机制进行,都观察到需要直系同源对的两个组分。此处研究的DSB修复反应让人联想到RecBCD/RecA反应,并提示了一种可能与其他系统(包括RAD52介导的重组)相关的通用机制。