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首页> 外文期刊>Applied Microbiology >The Small RNAs PA2952.1 and PrrH as Regulators of Virulence, Motility, and Iron Metabolism in Pseudomonas aeruginosa
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The Small RNAs PA2952.1 and PrrH as Regulators of Virulence, Motility, and Iron Metabolism in Pseudomonas aeruginosa

机译：小RNA PA2952.1和PRRH作为假鼠铜绿假单胞菌的毒力，运动和铁代谢调节剂

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Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen that undergoes swarming motility in response to semisolid conditions with amino acids as a nitrogen source. With a genome encoding hundreds of potential intergenic small RNAs (sRNAs), P. aeruginosa can easily adapt to different conditions and stresses. We previously identified 20 sRNAs that were differentially expressed (DE) under swarming conditions. Here, these sRNAs were overexpressed in strain PAO1 and were subjected to an array of phenotypic screens. Overexpression of the PrrH sRNA resulted in decreased swimming motility, whereas a Δ prrH mutant had decreased cytotoxicity and increased pyoverdine production. Overexpression of the previously uncharacterized PA2952.1 sRNA resulted in decreased swarming and swimming motilities, increased gentamicin and tobramycin resistance under swarming conditions, and increased trimethoprim susceptibility. Transcriptome sequencing (RNA-Seq) and proteomic analysis were performed on the wild type (WT) overexpressing PA2952.1 compared to the empty vector control under swarming conditions, and these revealed the differential expression (absolute fold change [FC] ≥ 1.5) of 784 genes and the differential abundance (absolute FC ≥ 1.25) of 59 proteins. Among these were found 73 transcriptional regulators, two-component systems, and sigma and anti-sigma factors. Downstream effectors included downregulated pilus and flagellar genes, the upregulated efflux pump MexGHI-OpmD, and the upregulated arn operon. Genes involved in iron and zinc uptake were generally upregulated, and certain pyoverdine genes were upregulated. Overall, the sRNAs PA2952.1 and PrrH appeared to be involved in regulating virulence-related programs in P. aeruginosa , including iron acquisition and motility.IMPORTANCE Due to the rising incidence of multidrug-resistant (MDR) strains and the difficulty of eliminating P. aeruginosa infections, it is important to understand the regulatory mechanisms that allow this bacterium to adapt to and thrive under a variety of conditions. Small RNAs (sRNAs) are one regulatory mechanism that allows bacteria to change the amount of protein synthesized. In this study, we overexpressed 20 different sRNAs in order to investigate how this might affect different bacterial behaviors. We found that one of the sRNAs, PrrH, played a role in swimming motility and virulence phenotypes, indicating a potentially important role in clinical infections. Another sRNA, PA2952.1, affected other clinically relevant phenotypes, including motility and antibiotic resistance. RNA-Seq and proteomics of the strain overexpressing PA2952.1 revealed the differential expression of 784 genes and 59 proteins, with a total of 73 regulatory factors. This substantial dysregulation indicates an important role for the sRNA PA2952.1.
机译：假单胞菌铜绿假单胞菌是一种革兰氏阴性机会主义病原体，响应氨基酸作为氮源的半固体条件经历蜂拥而至的运动。通过编码数百个潜在的基因组小RNA（SRNA），P.铜绿假单胞菌可以容易地适应不同的条件和应力。我们以前确定了20个SRNA，在蜂拥而至的条件下差异表达（DE）。这里，这些SRNA在应变PaO1中过表达，并进行了一系列表型筛网。 PRRH SRNA的过度表达导致游泳运动减少，而δPRRH突变体具有降低细胞毒性并增加百voverdine生产。预先表达先前无表达的PA2952.1SRNA导致蜂拥而至的蜂鸣和游泳能力下降，祖氨基和蜂鸣中的抗霉素抵抗力，并增加了甲肾上腺素敏感性。与蜂拥而至的条件下的空载体对照相比，在野生型（WT）过表达PA2952.1上进行转录体测序（RNA-SEQ）和蛋白质组学分析，这些差异表达（绝对折叠变化[FC]≥1.5） 784个基因和59个蛋白质的差异丰度（绝对Fc≥1.25）。其中有73个转录调节剂，双组分系统和Sigma和抗Σ因素。下游效果包括下调的菌落和鞭毛基因，上调的流出泵Mexghi-OPMD，以及上调的ARN操纵子。通常上调涉及铁和锌摄取的基因，并升高了某些百倍基因。总体而言，SRNA PA2952.1和PRRH似乎参与了调节铜绿假单胞菌的毒力相关的程序，包括铁采集和摩托斯。由于多药抗性（MDR）菌株的发病率上升和消除P的难度导致。铜绿假单胞菌感染，了解允许这种细菌适应和在各种条件下适应和茁壮成长的调节机制非常重要。小RNA（SRNA）是一种调节机制，允许细菌改变合成的蛋白质。在这项研究中，我们过表达了20种不同的SRNA，以研究这可能会影响不同的细菌行为。我们发现其中一个SRNA，PRRH在游泳运动和毒力表型中起作用，表明临床感染中的潜在重要作用。另一种SRNA，PA2952.1，影响了其他临床相关表型，包括运动性和抗生素抗性。 RNA-SEQ和过表达PA2952.1的菌株的蛋白质组学显示出784个基因和59个蛋白质的差异表达，共有73个调节因子。这种大量的失呼量表明SRNA PA2952.1的重要作用。

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