title: Potassium mediates Escherichia coli enzyme IIA(Ntr) -dependent regulation of sigma factor selectivity.
abstract: An Escherichia coli mutant devoid of enzyme IIA(Ntr) (EIIA(Ntr) ) of the nitrogen PTS is extremely sensitive to leucine-containing peptides due to decreased expression of acetohydroxy acid synthase. This decreased expression is due to defective potassium homeostasis. We further elucidate here the mechanism for regulation of gene expression by the intracellular level of K(+) . The leucine hypersensitivity of a ptsN (encoding EIIA(Ntr) ) mutant was suppressed by deleting rpoS, encoding the stationary phase σ factor. Despite intracellular levels of sigma factors comparable to the wild-type strain, most of the genes downregulated in a ptsN mutant are controlled by σ(70) , while all the upregulated genes are controlled by σ(S) , implying that the balance of sigma activities is modified by ptsN deletion. This change of sigma factor activities in the deletion mutant was found to be due to increased levels of K(+) . In vitro transcription assays demonstrated that a σ(70) controlled gene and a σ(S) controlled gene were differentially affected by potassium concentration. Biochemical studies revealed that K(+) is responsible for sigma factor competition by differentially influencing the binding of σ(70) and σ(S) to core RNA polymerase. Taken together, the data suggest that EIIA(Ntr) controls sigma factor selectivity by regulating the intracellular K(+) level.
authors.affiliation: Department of Biophysics and Chemical Biology, Seoul National University, Seoul 151-742, Republic of Korea.
authors.forename: Chang-Ro
authors.initials: CR
authors.lastname: Lee
authors.forename: Seung-Hyon
authors.initials: SH
authors.lastname: Cho
authors.forename: Hyun-Jin
authors.initials: HJ
authors.lastname: Kim
authors.forename: Miri
authors.initials: M
authors.lastname: Kim
authors.forename: Alan
authors.initials: A
authors.lastname: Peterkofsky
authors.forename: Yeong-Jae
authors.initials: YJ
authors.lastname: Seok
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000235:genetics / Q000378:metabolism*; D011188:Potassium / Q000378:metabolism*; D011485:Protein Binding; D012808:Sigma Factor / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D012808:Sigma Factor; C069567:sigma factor KatF protein, Bacteria; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C529959:PtsN protein, E coli; C081775:RNA polymerase sigma 70; D012321:DNA-Directed RNA Polymerases; D011188:Potassium