Present address: Department of Cancer Biology, College of Medicine & Life Sciences, The University of Toledo, Toledo OH 43614-1021, USA.
Department of Medical Microbiology & Immunology, and Program in Bioinformatics, College of Medicine & Life Sciences, The University of Toledo, Toledo OH 43614-1021, USA.
Microbiology (Reading). 2020 Nov;166(11):1047-1064. doi: 10.1099/mic.0.000980. Epub 2020 Oct 21.
Homologous recombination plays key roles in fundamental processes such as recovery from DNA damage and in bacterial horizontal gene transfer, yet there are still open questions about the distribution of recognized components of recombination machinery among bacteria and archaea. RecBCD helicase-nuclease plays a central role in recombination among Gammaproteobacteria like ; while bacteria in other phyla, like the Firmicute , use the related AddAB complex. The activity of at least some of these complexes is controlled by short DNA sequences called crossover hotspot instigator (Chi) sites. When RecBCD or AddAB complexes encounter an autologous Chi site during unwinding, they introduce a nick such that ssDNA with a free end is available to invade another duplex. If homologous DNA is present, RecA-dependent homologous recombination is promoted; if not (or if no autologous Chi site is present) the RecBCD/AddAB complex eventually degrades the DNA. We examined the distribution of and genes among bacteria, and sought ways to distinguish them unambiguously. We examined bacterial species among 33 phyla, finding some unexpected distribution patterns. and are less conserved than , with the orthologous and genes more conserved than the or genes. We were able to classify RecB vs. AddA and RecC vs. AddB in some bacteria where this had not previously been done. We used logo analysis to identify sequence segments that are conserved, but differ between the RecBC and AddAB proteins, to help future differentiation between members of these two families.
同源重组在 DNA 损伤恢复和细菌水平基因转移等基本过程中发挥着关键作用,但仍有一些关于重组机制的公认组件在细菌和古菌中的分布的问题尚未解决。RecBCD 解旋酶-核酸酶在 γ-变形菌(如 )的重组中起着核心作用;而在其他门的细菌中,如厚壁菌门 ,则使用相关的 AddAB 复合物。至少这些复合物中的一些的活性受到称为交叉热点引发(Chi)位点的短 DNA 序列的控制。当 RecBCD 或 AddAB 复合物在解旋过程中遇到同源 Chi 位点时,它们会引入一个切口,从而使带有游离末端的 ssDNA 可用于入侵另一个双链体。如果存在同源 DNA,则促进 RecA 依赖性同源重组;如果没有(或者没有同源 Chi 位点),RecBCD/AddAB 复合物最终会降解 DNA。我们检查了 和 基因在细菌中的分布,并寻求了明确区分它们的方法。我们检查了 33 个门的细菌物种,发现了一些意想不到的分布模式。 和 比 保守性差,而直系同源的 和 基因比 或 基因更保守。我们能够对以前没有做过的一些细菌中的 RecB 与 AddA 和 RecC 与 AddB 进行分类。我们使用 logo 分析来识别保守但在 RecBC 和 AddAB 蛋白之间存在差异的序列片段,以帮助未来区分这两个家族的成员。