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分析聚生型铜绿假单胞菌 lasR/rsaL 基因对的结构和表达模式揭示了它们之间的相互影响。

Analysis of the organization and expression patterns of the convergent Pseudomonas aeruginosa lasR/rsaL gene pair uncovers mutual influence.

机构信息

Department of Molecular Bacteriology, Helmholtz Centre for Infection Research, Braunschweig, Germany.

Institute of Molecular Bacteriology, TWINCORE Centre for Experimental and Clinical Infection Research, Hannover, Germany.

出版信息

Mol Microbiol. 2021 Apr;115(4):643-657. doi: 10.1111/mmi.14628. Epub 2020 Nov 11.

Abstract

The two adjacent genes encoding the major Pseudomonas aeruginosa quorum-sensing regulator, LasR, and its opponent, RsaL, overlap in their coding 3' ends and produce mRNA transcripts with long untranslated 3' ends that overlap with the sense transcripts of the gene on the opposing DNA strand. In this study, we evaluated whether the overlapping genes are involved in mutual regulatory events and studied interference by natural antisense transcripts. We introduced various gene expression constructs into a P. aeruginosa PA14 lasR/rsaL double deletion mutant, and found that although complementary RNA is produced, this does not interfere with the sense gene expression levels of lasR and rsaL and does not have functional consequences on down-stream gene regulation. Nevertheless, expression of lasR, but not of rsaL, was shown to be enhanced if transcription was terminated at the end of the respective gene so that no overlapping transcription was allowed. Our data indicate that the natural organization with a partial overlap at the 3' ends of the lasR/rsaL genes gives rise to a system of checks and balances to prevent dominant and unilateral control by LasR over the RsaL transcriptional regulator of opposing function.

摘要

编码铜绿假单胞菌主要群体感应调节因子 LasR 及其拮抗剂 RsaL 的两个相邻基因在其编码的 3' 末端重叠,并产生具有长非翻译 3' 末端的 mRNA 转录本,该转录本与相反 DNA 链上基因的有义转录本重叠。在这项研究中,我们评估了重叠基因是否参与相互调节事件,并研究了天然反义转录本的干扰。我们将各种基因表达构建体引入铜绿假单胞菌 PA14 lasR/rsaL 双缺失突变体中,发现尽管产生了互补 RNA,但这不会干扰 lasR 和 rsaL 的有意义基因表达水平,并且对下游基因调节没有功能后果。然而,如果转录在各自基因的末端终止,从而不允许重叠转录,则 lasR 的表达,但 rsaL 的表达不受影响。我们的数据表明,lasR/rsaL 基因 3' 末端的部分重叠的自然组织产生了一种制衡系统,以防止 LasR 对功能相反的 RsaL 转录调节剂进行主导和单方面控制。

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