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Alginate 和运动调节剂 AmrZ 对生物膜的分散响应调控至关重要。

The Alginate and Motility Regulator AmrZ is Essential for the Regulation of the Dispersion Response by Biofilms.

机构信息

Department of Biological Sciences, Binghamton Universitygrid.264260.4, Binghamton, New York, USA.

Binghamton Biofilm Research Center, Binghamton Universitygrid.264260.4, Binghamton, New York, USA.

出版信息

mSphere. 2022 Dec 21;7(6):e0050522. doi: 10.1128/msphere.00505-22. Epub 2022 Nov 14.

Abstract

Dispersion is an active process exhibited by Pseudomonas aeruginosa during the late stages of biofilm development or in response to various cues, including nitric oxide and glutamate. Upon cue sensing, biofilm cells employ enzymes that actively degrade the extracellular matrix, thereby allowing individual cells to become liberated. While the mechanism by which P. aeruginosa senses and relays dispersion cues has been characterized, little is known about how dispersion cue sensing mechanisms result in matrix degradation. Considering that the alginate and motility regulator AmrZ has been reported to regulate genes that play a role in dispersion, including those affecting virulence, c-di-GMP levels, Pel and Psl abundance, and motility, we asked whether AmrZ contributes to the regulation of dispersion. was found to be significantly increased in transcript abundance under dispersion-inducing conditions, with the inactivation of impairing dispersion by P. aeruginosa biofilms in response to glutamate and nitric oxide. While the overexpression of genes encoding matrix-degrading enzymes , , and/or resulted in the dispersion of wild-type biofilms, similar conditions failed to disperse biofilms formed by dt. Likewise, the inactivation of abrogated the hyperdispersive phenotype of PAO1/pJN-_G31A biofilms, with dt-impaired dispersion being independent of the expression, production, and activation of BdlA. Instead, dispersion was found to require the AmrZ-target genes and PA1891. Our findings indicate that AmrZ is essential for the regulation of dispersion by P. aeruginosa biofilms, functions downstream of BdlA postdispersion cue sensing, and regulates the expression of genes contributing to biofilm matrix degradation as well as and PA1891. In P. aeruginosa, biofilm dispersion has been well-characterized with respect to dispersion cue perception, matrix degradation, and the consequences of dispersion. While the intracellular signaling molecule c-di-GMP has been linked to many of the phenotypic changes ascribed to dispersion, including the modulation of motility and matrix production, little is known about the regulatory mechanisms leading to matrix degradation and cells actively leaving the biofilm. In this study, we report for the first time an essential role of the transcriptional regulator AmrZ and two AmrZ-dependent genes, and PA1891, in the dispersion response, thereby linking dispersion cue sensing via BdlA to the regulation of matrix degradation and to the ultimate liberation of bacterial cells from the biofilm.

摘要

分散是铜绿假单胞菌在生物膜发育后期或响应各种信号(包括一氧化氮和谷氨酸)时表现出的一种主动过程。在感知信号后,生物膜细胞利用酶主动降解细胞外基质,从而使单个细胞得以游离。虽然已经确定了铜绿假单胞菌感知和传递分散信号的机制,但对于分散信号感知机制如何导致基质降解知之甚少。鉴于已经报道了 Alginate 和运动调节剂 AmrZ 调节与分散相关的基因,包括影响毒力、c-di-GMP 水平、Pel 和 Psl 丰度以及运动性的基因,我们询问 AmrZ 是否有助于调节分散。发现其在分散诱导条件下的转录丰度显著增加,而谷氨酸和一氧化氮响应时, AmrZ 的失活会损害铜绿假单胞菌生物膜的分散。过表达编码基质降解酶的基因 、 、 和/或 导致野生型生物膜分散,但类似条件未能使 dt 形成的生物膜分散。同样, AmrZ 的失活消除了 PAO1/pJN-_G31A 生物膜的超分散表型,而 dt 受损的分散与 BdlA 的表达、产生和激活无关。相反,发现分散需要 AmrZ 靶基因 和 PA1891。我们的研究结果表明,AmrZ 是铜绿假单胞菌生物膜分散调节所必需的,在 BdlA 分散后信号感应中起作用,并调节参与生物膜基质降解以及 和 PA1891 的基因表达。在铜绿假单胞菌中,已经很好地描述了生物膜分散与分散信号感知、基质降解以及分散的后果有关。虽然细胞内信号分子 c-di-GMP 与归因于分散的许多表型变化有关,包括运动性和基质产生的调节,但对于导致基质降解和细菌细胞主动离开生物膜的调节机制知之甚少。在这项研究中,我们首次报道了转录调节剂 AmrZ 和两个 AmrZ 依赖性基因 和 PA1891 在分散反应中的重要作用,从而将通过 BdlA 进行的分散信号感知与基质降解的调节以及细菌细胞从生物膜中的最终释放联系起来。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/93d8/9769550/058c2ab21166/msphere.00505-22-f001.jpg

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