Key Laboratory of Chinese Ministry of Agriculture for Nuclear-Agricultural Sciences, Institute of Nuclear-Agricultural Sciences, Zhejiang University, Hangzhou, 310029, China.
Agriculture and Food Science School, Zhejiang Agriculture and Forestry University, Zhejiang, Lin'an, 311300, China.
Biochem Biophys Res Commun. 2019 Jun 4;513(3):740-745. doi: 10.1016/j.bbrc.2019.04.042. Epub 2019 Apr 13.
RecFOR and RecA are key recombination factors in Deinococcus radiodurans, a bacterium that possesses robust DNA repair capability and is also naturally transformable. While RecFOR functioning as a RecA loader during DNA repair has been established, their relative roles in transformation need further exploration. Here, we constructed recFOR and recA deletion mutants of D. radiodurans, and investigated the effect of these mutations on DNA transformation. recA deletion causes defects in both plasmid and chromosomal transformation. However, it was found that recFOR is not involved in chromosomal transformation, and that only recO and recR mutations compromise plasmid transformation. How recO, recR and recA mutations influence plasmid transformation was further examined by complementation plasmids. Interestingly, the transformation process remains defective in the recA mutant, but gets restored in the recO and recR mutants. This indicates that unlike RecA, RecOR may not be essential for DNA uptake. Therefore, we provide evidence that RecFOR is dispensable for RecA to protect incoming exogenous DNA and to catalyze recombination during transformation. Instead, RecO and RecR are likely to promote later steps in plasmid transformation.
RecFOR 和 RecA 是耐辐射球菌(Deinococcus radiodurans)中的关键重组因子,该菌具有强大的 DNA 修复能力,且天然可转化。虽然 RecFOR 在 DNA 修复过程中作为 RecA 加载器的功能已得到证实,但它们在转化中的相对作用仍需进一步探索。在此,我们构建了耐辐射球菌的 recFOR 和 recA 缺失突变体,并研究了这些突变对 DNA 转化的影响。recA 缺失导致质粒和染色体转化均出现缺陷。然而,研究发现 recFOR 不参与染色体转化,只有 recO 和 recR 突变会影响质粒转化。通过互补质粒进一步研究了 recO、recR 和 recA 突变如何影响质粒转化。有趣的是,recA 突变体中的转化过程仍然存在缺陷,但在 recO 和 recR 突变体中得到恢复。这表明,与 RecA 不同,RecOR 可能不是 DNA 摄取所必需的。因此,我们提供的证据表明,RecFOR 对于 RecA 保护进入的外源 DNA 并在转化过程中催化重组是可有可无的。相反,RecO 和 RecR 可能促进质粒转化的后期步骤。