Wardell Kayleigh, Haldenby Sam, Jones Nathan, Liddell Susan, Ngo Greg H P, Allers Thorsten
School of Life Sciences, University of Nottingham, Queen's Medical Centre, Nottingham, UK.
School of Biosciences, University of Nottingham, Sutton Bonington, UK.
DNA Repair (Amst). 2017 Jul;55:7-16. doi: 10.1016/j.dnarep.2017.04.005. Epub 2017 Apr 26.
Homologous recombination plays a central role in the repair of double-strand DNA breaks, the restart of stalled replication forks and the generation of genetic diversity. Regulation of recombination is essential since defects can lead to genome instability and chromosomal rearrangements. Strand exchange is a key step of recombination - it is catalysed by RecA in bacteria, Rad51/Dmc1 in eukaryotes and RadA in archaea. RadB, a paralogue of RadA, is present in many archaeal species. RadB has previously been proposed to function as a recombination mediator, assisting in RadA-mediated strand exchange. In this study, we use the archaeon Haloferax volcanii to provide evidence to support this hypothesis. We show that RadB is required for efficient recombination and survival following treatment with DNA-damaging agents, and we identify two point mutations in radA that suppress the ΔradB phenotype. Analysis of these point mutations leads us to propose that the role of RadB is to act as a recombination mediator, which it does by inducing a conformational change in RadA and thereby promoting its polymerisation on DNA.
同源重组在双链DNA断裂修复、停滞复制叉的重启以及遗传多样性的产生中起着核心作用。重组的调控至关重要,因为缺陷可能导致基因组不稳定和染色体重排。链交换是重组的关键步骤——在细菌中由RecA催化,在真核生物中由Rad51/Dmc1催化,在古细菌中由RadA催化。RadB是RadA的旁系同源物,存在于许多古细菌物种中。此前有人提出RadB作为重组介质发挥作用,协助RadA介导的链交换。在本研究中,我们利用嗜盐嗜碱菌提供证据支持这一假设。我们表明,在用DNA损伤剂处理后,RadB是高效重组和存活所必需的,并且我们在radA中鉴定出两个点突变,它们抑制了ΔradB表型。对这些点突变的分析使我们提出,RadB的作用是作为重组介质,它通过诱导RadA的构象变化,从而促进其在DNA上的聚合来实现这一作用。