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RecA-independent recombination: Dependence on the Escherichia coli RarA protein
Molecular Microbiology ( IF 3.6 ) Pub Date : 2020-11-28 , DOI: 10.1111/mmi.14655
Kanika Jain 1 , Elizabeth A Wood 1 , Zachary J Romero 1 , Michael M Cox 1
Affiliation  

Most, but not all, homologous genetic recombination in bacteria is mediated by the RecA recombinase. The mechanistic origin of RecA-independent recombination has remained enigmatic. Here, we demonstrate that the RarA protein makes a major enzymatic contribution to RecA-independent recombination. In particular, RarA makes substantial contributions to intermolecular recombination and to recombination events involving relatively short (<200 bp) homologous sequences, where RecA-mediated recombination is inefficient. The effects are seen here in plasmid-based recombination assays and in vivo cloning processes. Vestigial levels of recombination remain even when both RecA and RarA are absent. Additional pathways for RecA-independent recombination, possibly mediated by helicases, are suppressed by exonucleases ExoI and RecJ. Translesion DNA polymerases may also contribute. Our results provide additional substance to a previous report of a functional overlap between RecA and RarA.

中文翻译:

不依赖 RecA 的重组:依赖于大肠杆菌 RarA 蛋白

大多数(但不是全部)细菌中的同源基因重组是由 RecA 重组酶介导的。不依赖 RecA 的重组的机制起源仍然是个谜。在这里,我们证明 RarA 蛋白对不依赖于 RecA 的重组做出了主要的酶促贡献。特别是,RarA 对分子间重组和涉及相对较短(<200 bp)同源序列的重组事件做出了重大贡献,而 RecA 介导的重组效率较低。在基于质粒的重组测定和体内克隆过程中可以看到这种影响。即使 RecA 和 RarA 都不存在,重组的残余水平仍然存在。可能由解旋酶介导的不依赖于 RecA 的重组的其他途径被核酸外切酶 ExoI 和 RecJ 抑制。跨损伤 DNA 聚合酶也可能有所贡献。我们的结果为之前关于 RecA 和 RarA 之间功能重叠的报告提供了额外的实质内容。
更新日期:2020-11-28
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