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南极假单胞菌中响应氧气可用性和氧化应激的小 RNA。

Small RNAs in the Antarctic bacterium Pseudomonas extremaustralis responsive to oxygen availability and oxidative stress.

机构信息

IQUIBICEN-CONICET, Intendente Guiraldes 2160, 1428EGA, Buenos Aires, Argentina.

Institute of Molecular Infection Biology (IMIB), University of Würzburg, Würzburg, Germany.

出版信息

Environ Microbiol Rep. 2022 Aug;14(4):604-615. doi: 10.1111/1758-2229.13084. Epub 2022 Jun 10.

Abstract

Bacterial small non-coding RNAs (sRNAs) play key roles as genetic regulators, mediating in the adaptability to changing environmental conditions and stress responses. In this work, we analysed putative sRNAs identified by RNA-seq experiments in different aeration conditions in the extremophile bacterium P. extremaustralis. These analyses allowed the identification of 177 putative sRNAs under aerobiosis (A), microaerobiosis (M) and microaerobiosis after H O exposure (m-OS). The size and transcription profile of eight sRNAs with differential expression were verified by Northern blot. sRNA40, with unknown function but conserved in other Pseudomonas species, was selected to perform overexpression experiments followed by RNA-seq analysis. The overexpression of sRNA40 in P. extremaustralis resulted in significant expression changes of 19 genes with 14 differentially upregulated and five downregulated. Among the upregulated genes, eight transcripts corresponded to components of secretion systems, such as gspH, gspK, and gspM, belonging to the Type II secretion system, and rspO and rspP from Type III secretion system. Our results showed a novel sRNA which expression was triggered by low oxygen levels, and whose overexpression was associated with upregulation of selected components of protein secretion systems.

摘要

细菌小非编码 RNA(sRNA)作为遗传调节剂发挥着关键作用,介导对环境条件变化和应激反应的适应性。在这项工作中,我们分析了极端嗜热菌 P. extremaustralis 在不同通气条件下 RNA-seq 实验中鉴定出的假定 sRNA。这些分析鉴定了在需氧条件(A)、微需氧条件(M)和 H2O2 暴露后的微需氧条件(m-OS)下的 177 个假定 sRNA。通过 Northern blot 验证了 8 个具有差异表达的 sRNA 的大小和转录谱。sRNA40 具有未知功能,但在其他假单胞菌物种中保守,选择其进行过表达实验,然后进行 RNA-seq 分析。sRNA40 在 P. extremaustralis 中的过表达导致 19 个基因的显著表达变化,其中 14 个上调,5 个下调。在上调的基因中,有 8 个转录本对应于分泌系统的组成部分,如 gspH、gspK 和 gspM,属于 II 型分泌系统,以及 Type III 分泌系统的 rspO 和 rspP。我们的结果显示了一种新的 sRNA,其表达受到低氧水平的触发,其过表达与选定的蛋白分泌系统组成部分的上调有关。

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