Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
Int J Mol Sci. 2020 Apr 30;21(9):3200. doi: 10.3390/ijms21093200.
is a polyextremophilic bacterium well known for its extreme resistance to irradiation, oxidative stress, and other damaging conditions. Many small noncoding RNAs (ncRNAs) in have been identified by deep sequencing analysis and computational predictions. However, the precise roles of ncRNAs and their target genes in the oxidative stress response have not been investigated. Here, we report the identification and characterization of a novel ncRNA named OsiR (for oxidative stress-induced ncRNA). Oxidative stress tolerance analysis showed that deleting osiR significantly decreased viability, total antioxidant capacity, and catalase activity in D. radiodurans under oxidative stress conditions. Comparative phenotypic and qRT-PCR analyses of an osiR mutant identify a role of OsiR in regulating the expression of the catalase gene katE2. Microscale thermophoresis and genetic complementation showed that a 21-nt sequence in the stem-loop structure of OsiR (204-244 nt) directly base pairs with its counterpart in the coding region of katE2 mRNA (843-866 nt) via a 19 nt region. In addition, deletion of katE2 caused a significant reduction of catalase activity and oxidative stress tolerance similar to that observed in an mutant. Our results show that OsiR positively regulates oxidative stress tolerance in by increasing the mRNA stability and translation efficiency of . This work provides a new regulatory pathway mediated by ncRNA for the oxidative stress response that most likely contributes to the extreme tolerances of
是一种多极端环境细菌,以其对辐射、氧化应激和其他损伤条件的极端抗性而闻名。通过深度测序分析和计算预测,已经在 中鉴定出许多小非编码 RNA(ncRNA)。然而,ncRNA 及其靶基因在氧化应激反应中的精确作用尚未得到研究。在这里,我们报告了一种新型 ncRNA 的鉴定和表征,命名为 OsiR(用于氧化应激诱导的 ncRNA)。氧化应激耐受性分析表明,在氧化应激条件下,缺失 osiR 会显著降低 D. radiodurans 的存活率、总抗氧化能力和过氧化氢酶活性。osiR 突变体的表型和 qRT-PCR 分析比较确定了 OsiR 在调节过氧化氢酶基因 katE2 表达中的作用。微尺度热泳和遗传互补表明,OsiR 茎环结构中的 21 个核苷酸序列(204-244 个核苷酸)通过 19 个核苷酸区域与 katE2 mRNA 编码区的互补序列(843-866 个核苷酸)直接碱基配对。此外,katE2 的缺失导致过氧化氢酶活性和氧化应激耐受性的显著降低,类似于在 突变体中观察到的情况。我们的结果表明,OsiR 通过增加 的 mRNA 稳定性和翻译效率,正向调节 在氧化应激中的耐受性。这项工作提供了一种新的由 ncRNA 介导的氧化应激反应调控途径,这很可能有助于其极端耐受性。