PqsE adapts the activity of the Pseudomonas aeruginosa quorum-sensing transcription factor RhlR to both autoinducer concentration and promoter sequence identity.

IF 2.7 3区 生物学 Q3 MICROBIOLOGY
Journal of Bacteriology Pub Date : 2025-05-22 Epub Date: 2025-04-17 DOI:10.1128/jb.00516-24
Bilalay V Tchadi, Jesse J Derringer, Anna K Detweiler, Isabelle R Taylor
{"title":"PqsE adapts the activity of the <i>Pseudomonas aeruginosa</i> quorum-sensing transcription factor RhlR to both autoinducer concentration and promoter sequence identity.","authors":"Bilalay V Tchadi, Jesse J Derringer, Anna K Detweiler, Isabelle R Taylor","doi":"10.1128/jb.00516-24","DOIUrl":null,"url":null,"abstract":"<p><p><i>Pseudomonas aeruginosa</i> is an opportunistic human pathogen that poses a significant health threat. Many pathogenic behaviors of <i>P. aeruginosa</i> are under control of the bacterial cell-cell communication system known as quorum sensing (QS). One of the QS master regulators, RhlR, is a receptor/transcription factor that not only relies on binding of its canonical ligand, <i>N</i>-butyrylhomoserine lactone (C4-HSL), but additionally requires a protein-protein interaction with the enzyme, PqsE. We constructed heterologous reporter strains in <i>Escherichia coli</i> that allow measurements of the reliance of RhlR on C4-HSL and/or PqsE binding for the ability to activate transcription of three RhlR-regulated genes: <i>rhlA</i> (PqsE independent), <i>phzM</i> (PqsE dependent), and <i>azeB</i> (PqsE inhibited). Analogous assays measuring activation of the three genes in <i>P. aeruginosa</i> were performed, and the patterns observed correlated tightly with the heterologous reporter assays. These results confirm that the binding of PqsE to RhlR is able to fine-tune RhlR transcription factor activity in a promoter-specific manner and prove that this ability is independent of other factors present in <i>P. aeruginosa</i>.IMPORTANCE<i>Pseudomonas aeruginosa</i> is an opportunistic human pathogen that can cause fatal infections. There exists an urgent need for new, effective antimicrobial agents to combat <i>P. aeruginosa</i>. The PqsE-RhlR protein-protein interaction is essential for <i>P. aeruginosa</i> to be able to make toxins, form biofilms, and infect host organisms. In this study, we use both non-native models in <i>Escherichia coli</i> and measurements of gene expression/toxin production in <i>P. aeruginosa</i> to show that the PqsE-RhlR interaction enables fine-tuned gene expression and a heightened ability of <i>P. aeruginosa</i> to adapt to external conditions. These findings will be highly valuable as continued efforts are made to design inhibitors of the PqsE-RhlR interaction and test them as potential antimicrobial agents against <i>P. aeruginosa</i> infections.</p>","PeriodicalId":15107,"journal":{"name":"Journal of Bacteriology","volume":" ","pages":"e0051624"},"PeriodicalIF":2.7000,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12096825/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Bacteriology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1128/jb.00516-24","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/4/17 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

Pseudomonas aeruginosa is an opportunistic human pathogen that poses a significant health threat. Many pathogenic behaviors of P. aeruginosa are under control of the bacterial cell-cell communication system known as quorum sensing (QS). One of the QS master regulators, RhlR, is a receptor/transcription factor that not only relies on binding of its canonical ligand, N-butyrylhomoserine lactone (C4-HSL), but additionally requires a protein-protein interaction with the enzyme, PqsE. We constructed heterologous reporter strains in Escherichia coli that allow measurements of the reliance of RhlR on C4-HSL and/or PqsE binding for the ability to activate transcription of three RhlR-regulated genes: rhlA (PqsE independent), phzM (PqsE dependent), and azeB (PqsE inhibited). Analogous assays measuring activation of the three genes in P. aeruginosa were performed, and the patterns observed correlated tightly with the heterologous reporter assays. These results confirm that the binding of PqsE to RhlR is able to fine-tune RhlR transcription factor activity in a promoter-specific manner and prove that this ability is independent of other factors present in P. aeruginosa.IMPORTANCEPseudomonas aeruginosa is an opportunistic human pathogen that can cause fatal infections. There exists an urgent need for new, effective antimicrobial agents to combat P. aeruginosa. The PqsE-RhlR protein-protein interaction is essential for P. aeruginosa to be able to make toxins, form biofilms, and infect host organisms. In this study, we use both non-native models in Escherichia coli and measurements of gene expression/toxin production in P. aeruginosa to show that the PqsE-RhlR interaction enables fine-tuned gene expression and a heightened ability of P. aeruginosa to adapt to external conditions. These findings will be highly valuable as continued efforts are made to design inhibitors of the PqsE-RhlR interaction and test them as potential antimicrobial agents against P. aeruginosa infections.

PqsE使铜绿假单胞菌群体感应转录因子RhlR的活性适应于自诱导剂浓度和启动子序列的一致性。
铜绿假单胞菌是一种机会性的人类病原体,对健康构成重大威胁。铜绿假单胞菌的许多致病行为是由细菌细胞间通讯系统控制的,称为群体感应(quorum sensing, QS)。QS主调控因子之一RhlR是一种受体/转录因子,它不仅依赖于其典型配体n-丁基同丝氨酸内酯(C4-HSL)的结合,还需要与酶PqsE进行蛋白-蛋白相互作用。我们在大肠杆菌中构建了异源报告菌株,可以测量RhlR对C4-HSL和/或PqsE结合的依赖,以激活三种RhlR调节基因的转录能力:rhlA (PqsE独立),phzM (PqsE依赖)和azeB (PqsE抑制)。在铜绿假单胞菌中进行了测量这三个基因激活的类似分析,观察到的模式与异种报告基因的分析密切相关。这些结果证实,PqsE与RhlR结合能够以启动子特异性的方式微调RhlR转录因子的活性,并证明这种能力独立于铜绿假单胞菌中存在的其他因子。铜绿假单胞菌是一种机会性的人类病原体,可引起致命的感染。目前迫切需要新的、有效的抗菌药物来对抗铜绿假单胞菌。PqsE-RhlR蛋白-蛋白相互作用是铜绿假单胞菌制造毒素、形成生物膜和感染宿主生物所必需的。在这项研究中,我们使用了大肠杆菌的非天然模型和铜绿假单胞菌的基因表达/毒素产生的测量,表明PqsE-RhlR相互作用可以微调基因表达,提高铜绿假单胞菌适应外部条件的能力。这些发现对于设计PqsE-RhlR相互作用抑制剂并测试其作为铜绿假单胞菌感染的潜在抗菌药物具有很高的价值。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Journal of Bacteriology
Journal of Bacteriology 生物-微生物学
CiteScore
6.10
自引率
9.40%
发文量
324
审稿时长
1.3 months
期刊介绍: The Journal of Bacteriology (JB) publishes research articles that probe fundamental processes in bacteria, archaea and their viruses, and the molecular mechanisms by which they interact with each other and with their hosts and their environments.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信