Bilalay V Tchadi, Jesse J Derringer, Anna K Detweiler, Isabelle R Taylor
{"title":"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。","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":"{\"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}","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}
PqsE adapts the activity of the Pseudomonas aeruginosa quorum-sensing transcription factor RhlR to both autoinducer concentration and promoter sequence identity.
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.
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.