Knockouts of RecA-Like Proteins RadC1 and RadC2 Have Distinct Responses to DNA Damage Agents in Sulfolobus islandicus
Knockouts of RecA-Like Proteins RadC1 and RadC2 Have Distinct Responses to DNA Damage Agents in Sulfolobus islandicus
复制标题
RecA 样蛋白 RadC1 和 RadC2 的敲除对岛硫化叶菌中的 DNA 损伤剂有不同的反应
DOI：
10.1016/j.jgg.2013.05.004
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发表时间：
2013-10-01
影响因子：
5.9
通讯作者：
Shen, Yu-Long
中科院分区：
文献类型：
--
作者：
Liang, Peng-Juan;Han, Wen-Yuan;Shen, Yu-Long
RecA家族重组酶在维持基因组完整性方面发挥重要作用。一组名为RadC的RecA样蛋白存在于所有古细菌中，但其体内功能尚不清楚。在这项研究中，我们进行了系统发育和遗传分析的两个RadC蛋白从硫化叶菌islandicus。与重组酶（RecA、RadA和Rad51）和重组酶旁系同源物（例如，RadB、Rad55和Rad51B）。使用最近建立的S. islandicus遗传系统中，构建了radC1和radC2的缺失和过表达菌株。缺失的radC1使细胞更敏感的DNA损伤剂，甲基甲磺酸酯（MMS），羟基脲（HU），和紫外线（UV）辐射，比野生型，和一个三角洲radC1三角洲radC2双缺失菌株更敏感的顺铂和MMS比三角洲radC1单缺失突变体。此外，异位表达His标记的RadC1揭示，RadC1与推定的结构特异性核酸酶和ATP酶，这是高度保守的古细菌共纯化。我们的研究结果表明，RadC1和RadC2都参与DNA修复。RadC1可能在DNA修复中发挥一般或主要作用，而RadC2在响应特定DNA损伤的DNA修复中发挥作用。
RecA family recombinases play essential roles in maintaining genome integrity. A group of RecA-like proteins named RadC are present in all archaea, but their in vivo functions remain unclear. In this study, we performed phylogenetic and genetic analysis of two RadC proteins from Sulfolobus islandicus. RadC is closer to the KaiC lineage of cyanobacteria and proteobacteria than to the lineage of the recombinases (RecA, RadA, and Rad51) and the recombinase paralogs (e.g., RadB, Rad55, and Rad51B). Using the recently-stablished S. islandicus genetic system, we constructed deletion and over-expression strains of radC1 and radC2. Deletion of radC1 rendered the cells more sensitive to DNA damaging agents, methyl methanesulfonate (MMS), hydroxyurea (HU), and ultraviolet (UV) radiation, than the wild type, and a Delta radC1 Delta radC2 double deletion strain was more sensitive to cisplatin and MMS than the Delta radC1 single deletion mutant. In addition, ectopic expression of His-tagged RadC1 revealed that RadC1 was co-purified with a putative structure-specific nuclease and ATPase, which is highly conserved in archaea. Our results indicate that both RadC1 and RadC2 are involved in DNA repair. RadC1 may play a general or primary role in DNA repair, while RadC2 plays a role in DNA repair in response to specific DNA damages.