Key Laboratory for Nuclear-Agricultural Sciences of Chinese Ministry of Agriculture and Zhejiang Province, Institute of Nuclear-Agricultural Sciences, Zhejiang University, Hangzhou, P. R. China.
J Microbiol. 2010 Aug;48(4):473-9. doi: 10.1007/s12275-010-0043-8. Epub 2010 Aug 20.
A novel OxyR (DR0615) with one conserved cysteine that senses hydrogen peroxide in Deinococcus radiodurans had been identified in our previous work. Comparative genomics revealed that D. radiodurans possesses another OxyR homolog, OxyR(2) (DRA0336). In this study, we constructed the deletion mutant of oxyR(2) and the double mutant of both the OxyR homologs to investigate the role of OxyR in response to oxidative stress in D. Radiodurans. Deletion of oxyR(2) resulted in an obviously increased sensitivity to hydrogen peroxide, and the double mutant for oxyR and oxyR(2) was significantly more sensitive than any of the two single mutants. The total catalase activity of the double mutant was lower than that of any of the single mutants, and reactive oxygen species (ROS) accumulated to a greater extent. DNA microarray analysis further suggested that oxyR(2) was involved in antioxidation mechanisms. Site-direct mutagenesis and complementation analysis revealed that C(228) in OxyR(2) was essential. This is the first report of the presence of two OxyR in one organism. These results suggest that D. radiodurans OxyR and OxyR(2) function together to protect the cell against oxidative stress.
在我们之前的工作中,已经鉴定出一种新型的 OxyR(DR0615),它在耐辐射球菌中含有一个保守的半胱氨酸,能够感应过氧化氢。比较基因组学表明,耐辐射球菌还拥有另一个 OxyR 同源物,OxyR(2)(DRA0336)。在这项研究中,我们构建了 oxyR(2)的缺失突变体和两个 OxyR 同源物的双突变体,以研究 OxyR 在耐辐射球菌应对氧化应激中的作用。oxyR(2)的缺失导致对过氧化氢的敏感性明显增加,而 oxyR 和 oxyR(2)的双突变体比任何两个单突变体都更敏感。双突变体的总过氧化氢酶活性低于任何一个单突变体,并且活性氧(ROS)积累到更大程度。DNA 微阵列分析进一步表明,oxyR(2)参与了抗氧化机制。定点突变和互补分析表明,OxyR(2)中的 C(228)是必需的。这是在一个生物体中存在两个 OxyR 的首次报道。这些结果表明,耐辐射球菌的 OxyR 和 OxyR(2)共同作用以保护细胞免受氧化应激。