[Regulation of pyocyanin biosynthesis by transcriptional factor sigma38 in Pseudomonas aeruginosa PAO1].

微生物学报 Pub Date : 2017-02-04
Jing Miao, Xiaoyan Chi, Yanhua Wang, Zhibin Feng, Wenwen Xue, Run Huang, Haoyi Zhang, Lingqian Tian, Hongqian Zhang, Junjie Zhai, Yihe Ge
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Abstract

Pyocyanin, an important virulence factor, is synthesized and secreted by Pseudomonas aeruginosa PAO1and plays a critical role in pathogen-host interaction during infection. Sigma38 (σ38, σS) is a central regulator for many virulence production in pathogens.

Objective: Our aim is to identify expression and regulation of two phenazine-producing operons mediated by the sigma38 factor in Pseudomonas aeruginosa PAO1.

Methods: We first cloned the flanking fragments of rpoS from the chromosomal DNA of P. aeruginosa PAO1 and constructed the deletion mutant ΔrpoS with the insertion of gentamycin resistance cassette (aacC1). Complementation of rpoS was then carried out after construction and introduction of pME10S (containing the whole rpoS region). Finally, we created the mutant ΔrpoSphz1 and ΔrpoSphz2, and measured pyocyanin production by these mutants in GA medium, using the parental strain Δphz1 and Δphz2 as controls.

Results: In GA medium, pyocyanin production by mutant ΔrpoS increased dramatically in comparison with the wild-type strain PAO1. Production of pyocyanin, however, was decreased to the level of the wild-type strain with complementation of the derivative ΔrpoS harboring pME10S. Mutant ΔrpoSphz2 produced much more pyocyanin than mutant Δphz2. Mutant ΔrpoSphz1, however, produced much less pyocyanin than mutant Δphz1.

Conclusion: By positively regulating the expression of phz2 and negatively regulating the phz1, sigma38 factor exerts negative modulation on pyocyanin biosynthesis in P. aeruginosa PAO1.

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[铜绿假单胞菌PAO1中转录因子sigma38对pyocyanin生物合成的调控]。
Pyocyanin是一种重要的毒力因子,由铜绿假单胞菌pao1合成并分泌,在感染过程中病原菌与宿主相互作用中起关键作用。Sigma38 (σ38, σS)是病原菌中许多毒力产生的中心调控因子。目的:研究铜绿假单胞菌PAO1中sigma38因子介导的两个产非那嗪操纵子的表达和调控。方法:首先从铜绿假单胞菌PAO1染色体DNA中克隆rpoS的侧翼片段,构建插入庆大霉素耐药盒(aacC1)的缺失突变体ΔrpoS。在构建并引入pME10S(包含整个rpoS区域)后,对rpoS进行互补。最后,我们创建了突变体ΔrpoSphz1和ΔrpoSphz2,并以亲本菌株Δphz1和Δphz2为对照,在GA培养基中测量了这些突变体的pyocyanin产量。结果:在GA培养基中,与野生型菌株PAO1相比,突变体ΔrpoS的pyocyanin产量显著增加。然而,与含有pME10S的衍生物ΔrpoS互补后,pyocyanin的产量下降到野生型菌株的水平。突变体ΔrpoSphz2比突变体Δphz2产生更多的pyocyanin。然而,突变体ΔrpoSphz1产生的pyocyanin比突变体Δphz1少得多。结论:sigma38因子通过正向调控phz2的表达,负向调控phz1的表达,对P. aeruginosa PAO1中花青素的合成起负向调控作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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期刊介绍: Acta Microbiologica Sinica(AMS) is a peer-reviewed monthly (one volume per year)international journal,founded in 1953.It covers a wide range of topics in the areas of general and applied microbiology.The journal publishes original papers,reviews in microbiological science,and short communications describing unusual observations. Acta Microbiologica Sinica has been indexed in Index Copernicus (IC),Chemical Abstract (CA),Excerpt Medica Database (EMBASE),AJ of Viniti (Russia),Biological Abstracts (BA),Chinese Science Citation Database (CSCD),China National Knowledge Infrastructure(CNKI),Institute of Scientific and Technical Information of China(ISTIC),Chinese Journal Citation Report(CJCR),Chinese Biological Abstracts,Chinese Pharmaceutical Abstracts,Chinese Medical Abstracts and Chinese Science Abstracts.
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