Department of Molecular Biology, Ruđer Bošković Institute, Bijenička 54, 10000 Zagreb, Croatia.
Res Microbiol. 2013 Dec;164(10):987-97. doi: 10.1016/j.resmic.2013.08.008. Epub 2013 Sep 12.
The RecQ helicase is required by the RecF recombination pathway that is operative in recBC(D) sbcB sbcC(D) mutants of Escherichia coli. Genetic data suggest that RecQ participates in resection of DNA ends during initiation of recombination. In vitro, RecQ can unwind a variety of DNA substrates, including recombination intermediates such as D-loops and Holliday junctions. However, its potential role in processing of recombination intermediates during the late stage of the RecF pathway has not been genetically tested. Here we studied the effect of a recQ mutation on transductional recombination and DNA repair after γ-irradiation in ΔrecBCD ΔsbcB sbcC strains deficient for RuvABC, RecG and XerC proteins. RuvABC and RecG proteins process recombination intermediates in the late stage of recombination, whereas XerC is required to resolve chromosome dimers formed upon recombination. Our results do not reveal any substantial synergistic effect between the recQ mutation, on one hand, and ruvABC, recG and xerC mutations on the other. In addition, the recQ mutation suppresses chromosome segregation defects in γ-irradiated ruvABC recG and xerC mutants. These results suggest that RecQ acts upstream of RuvABC, RecG and XerC proteins, a finding that is compatible with its primary role in initiation of the RecF recombination pathway.
RecQ 解旋酶是 RecF 重组途径所必需的,该途径在大肠杆菌 recBC(D) sbcB sbcC(D) 突变体中起作用。遗传数据表明,RecQ 参与了重组起始时 DNA 末端的切除。在体外,RecQ 可以解开多种 DNA 底物,包括 D 环和 Holliday 连接等重组中间体。然而,它在 RecF 途径晚期处理重组中间体方面的潜在作用尚未通过遗传测试进行验证。在这里,我们研究了 recQ 突变对 RuvABC、RecG 和 XerC 蛋白缺失的 ΔrecBCD ΔsbcB sbcC 菌株中 γ 射线诱导的转导性重组和 DNA 修复的影响。RuvABC 和 RecG 蛋白在重组后期处理重组中间体,而 XerC 则需要解决重组形成的染色体二聚体。我们的结果没有揭示 recQ 突变与 ruvABC、recG 和 xerC 突变之间的任何实质性协同作用。此外,recQ 突变抑制了 γ 射线照射的 ruvABC recG 和 xerC 突变体中的染色体分离缺陷。这些结果表明 RecQ 在上游作用于 RuvABC、RecG 和 XerC 蛋白,这与它在 RecF 重组途径起始中的主要作用是一致的。