Characterization of mutations in the PAS domain of the EvgS sensor kinase selected by laboratory evolution for acid resistance in Escherichia coli. Issue 5 (24th July 2014)

• Record Type: Journal Article
• Authors: Johnson, Matthew D.; Bell, James; Clarke, Kim; Chandler, Rachel; Pathak, Prachi; Xia, Yandong; Marshall, Robert L.; Weinstock, George M.; Loman, Nicholas J.; Winn, Peter J.; Lund, Peter A.
• Is Part Of: Molecular microbiology. Volume 93:Issue 5(2014)
• Page Start: 911
• Page End: 927
• Publication Date: 2014-07-24
• Subjects: Molecular microbiology -- Periodicals; 572.829
• DOI: 10.1111/mmi.12704
• Languages: English
• ISSNs: 0950-382X

Abstract

<abstract abstract-type="main"> <title>Summary</title> <p>Laboratory‐based evolution and whole‐genome sequencing can link genotype and phenotype. We used evolution of acid resistance in exponential phase <italic>E</italic><italic>scherichia coli</italic> to study resistance to a lethal stress. Iterative selection at pH 2.5 generated five populations that were resistant to low pH in early exponential phase. Genome sequencing revealed multiple mutations, but the only gene mutated in all strains was <italic>evgS</italic>, part of a two‐component system that has already been implicated in acid resistance. All these mutations were in the cytoplasmic PAS domain of EvgS, and were shown to be solely responsible for the resistant phenotype, causing strong upregulation at neutral pH of genes normally induced by low pH. Resistance to pH 2.5 in these strains did not require the transporter GadC, or the sigma factor RpoS. We found that EvgS‐dependent constitutive acid resistance to pH 2.5 was retained in the absence of the regulators GadE or YdeO, but was lost if the oxidoreductase YdeP was also absent. A deletion in the periplasmic domain of EvgS abolished the response to low pH, but not the activity of the constitutive mutants. On the basis of these results we propose a model for how EvgS may become activated by low pH.</p> </abstract>