Institute of Experimental Botany, Czech Academy of Sciences, v.v.i., Na Karlovce 1, 16000 Prague 6, Czech Republic.
Institute of Experimental Botany, Czech Academy of Sciences, v.v.i., Rozvojová 263, 16502 Prague 6, Czech Republic.
Genes (Basel). 2023 Jan 24;14(2):305. doi: 10.3390/genes14020305.
RAD51 is involved in finding and invading homologous DNA sequences for accurate homologous recombination (HR). Its paralogs have evolved to regulate and promote RAD51 functions. The efficient gene targeting and high HR rates are unique in plants only in the moss (). In addition to two functionally equivalent genes ( and ), other paralogues were also identified in . For elucidation of RAD51's involvement during DSB repair, two knockout lines were constructed, one mutated in both genes () and the second with mutated gene (). Both lines are equally hypersensitive to bleomycin, in contrast to their very different DSB repair efficiency. Whereas DSB repair in is even faster than in WT, in , it is slow, particularly during the second phase of repair kinetic. We interpret these results as and - being true functional homologs of ancestral involved in the homology search during HR. Absence of RAD51 redirects DSB repair to the fast NHEJ pathway and leads to a reduced 5S and 18S rDNA copy number. The exact role of the RAD51B paralog remains unclear, though it is important in damage recognition and orchestrating HR response.
RAD51 参与寻找和入侵同源 DNA 序列以实现准确的同源重组 (HR)。其同源物进化为调节和促进 RAD51 的功能。有效的基因靶向和高 HR 率是植物所特有的,在苔藓中仅发现 ()。除了两个功能等效的基因(和)之外,还在 中鉴定到其他 同源物。为了阐明 RAD51 在 DSB 修复过程中的作用,构建了两个敲除系,一个在两个 基因()中发生突变,另一个在 基因()中发生突变。这两个系对博来霉素均表现出同等的超敏性,与它们在 DSB 修复效率上的差异形成鲜明对比。尽管在 中 DSB 修复甚至比 WT 更快,但在 中,它很慢,尤其是在修复动力学的第二阶段。我们将这些结果解释为 和 是参与 HR 同源搜索的祖先 的真正功能同源物。RAD51 的缺失将 DSB 修复重定向到快速的 NHEJ 途径,并导致 5S 和 18S rDNA 拷贝数减少。RAD51B 同源物的确切作用仍不清楚,但它在损伤识别和协调 HR 反应中很重要。