PMID:11985727

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Citation

McLaggan, D, Jones, MA, Gouesbet, G, Levina, N, Lindey, S, Epstein, W and Booth, IR (2002) Analysis of the kefA2 mutation suggests that KefA is a cation-specific channel involved in osmotic adaptation in Escherichia coli. Mol. Microbiol. 43:521-36

Abstract

Mechanosensitive channels play an essential role in the regulation of turgor pressure in bacteria. In Escherichia coli, there are multiple mechanosensitive channels that have been characterized genetically: MscL, YggB and KefA. In this report, we describe the cloning of the kefA gene, the organization of the KefA protein and the phenotype of a missense mutation, kefA, which affects the KefA mechanosensitive channel. The altered function of the channel is manifest through increased sensitivity to K+ during growth at low osmolarity and complete inhibition of growth in media containing high K+ concentrations (0.6 M) in the presence of betaine or proline. Growth in high Na+ medium (0.6 M NaCl plus 20 mM K+) is normal. Analysis of the cytoplasmic pools shows that the mutant cannot regulate the K+ content of the cytoplasm when grown in high K+ medium. However, regulation of pools of amino acids is essentially normal and the mutant can accumulate high pools of proline during growth inhibition. The mutant shows increased sensitivity to acid hypo-osmotic shock (transition from neutral to acid pH combined with a reduction in osmolarity). The data are consistent with abnormal regulation of KefA in the presence of high K+ concentrations and either betaine or proline.

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Keywords

Acids; Adaptation, Physiological; Bacterial Proteins/genetics; Bacterial Proteins/physiology; Cations; Escherichia coli/genetics; Escherichia coli/physiology; Escherichia coli Proteins/physiology; Genes, Bacterial; Glutamic Acid/metabolism; Mutagenesis; Osmolar Concentration; Osmosis; Potassium/metabolism; Potassium Channels/genetics; Potassium Channels/physiology; Proline; Trehalose/biosynthesis; Water-Electrolyte Balance/physiology

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