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We previously discovered that a mutation in the triosephosphate isomerase gene, <jats:italic>tpiA<\/jats:italic>, in <jats:italic>Pseudomonas aeruginosa<\/jats:italic> led to defective type III secretion and increased susceptibility to aminoglycoside antibiotics. In this study, we found that the <jats:italic>tpiA<\/jats:italic> mutation enhances the Las quorum sensing system due to reduced translation of the negative regulator RsaL. Further investigations demonstrated an upregulation of CspC, a CspA family protein that represses <jats:italic>rsaL<\/jats:italic> translation. DNA pull-down assay, along with genetic studies, revealed the role of AgtR in regulating <jats:italic>cspC<\/jats:italic> transcription. AgtR is known to regulate pyocyanin production in response to N-acetylglucosamine (GlcNAc), contributing to competition against <jats:italic>Staphylococcus aureus<\/jats:italic>. We demonstrated that CspC activates the Las quorum sensing system and subsequent pyocyanin production in response to GlcNAc and <jats:italic>S. aureus<\/jats:italic>. Overall, our results elucidate the AgtR-CspC-RsaL-LasI pathway that regulates bacterial virulence factors and its role in competition against <jats:italic>S. aureus<\/jats:italic>.<\/jats:p>","DOI":"10.1371\/journal.ppat.1013054","type":"journal-article","created":{"date-parts":[[2025,4,8]],"date-time":"2025-04-08T18:19:27Z","timestamp":1744136367000},"page":"e1013054","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":5,"title":["Identification of the Pseudomonas aeruginosa AgtR-CspC-RsaL pathway that controls Las quorum sensing in response to metabolic perturbation and Staphylococcus 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