A single amino acid substitution in the C terminus of OmpR alters DNA recognition and phosphorylation

Van K. Tran, Ricardo Oropeza, Linda Kenney

Research output: Contribution to journalArticle

46 Scopus citations

Abstract

In bacteria and lower eukaryotes, adaptation to changes in the environment is often mediated by two-component regulatory systems. Such systems provide the basis for chemotaxis, nitrogen and phosphate regulation and adaptation to osmotic stress, for example. In Escherichia coli, the sensor kinase EnvZ detects a change in the osmotic environment and phosphorylates the response regulator OmpR. Phospho-OmpR binds to the regulatory regions of the porin genes ompF and ompC, and alters their expression. Recent evidence suggests that OmpR functions as a global regulator, regulating additional genes besides the porin genes. In this study, we have characterized a previously isolated OmpR2 mutant (V203M) that constitutively activates ompF and fails to express ompC. Because the substitution was located in the C-terminal DNA-binding domain, it had been assumed that the substitution would not affect phosphorylation of the N-terminal domain of OmpR. Our results indicate that this substitution completely eliminates phosphorylation by a small phosphate donor, acetyl phosphate, but not phosphorylation by the kinase EnvZ. The mutant OmpR has altered dephosphorylation kinetics and altered binding affinities to both ompF and ompC sites compared to the wild-type. Thus, a single amino acid substitution in the C-terminal DNA-binding domain has dramatic effects on the N-terminal phosphorylation domain. Most strikingly, we have identified a single base change in the OmpR binding site of ompC that restores high-affinity binding activity by the mutant. We interpret our results in the context of a model for porin gene expression. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)1257-1270
Number of pages14
JournalJournal of Molecular Biology
Volume299
Issue number5
DOIs
StatePublished - Jun 23 2000
Externally publishedYes

Keywords

  • Osmoregulation
  • Response regulator
  • Signal transduction
  • Two-component regulatory system
  • Winged helix-turn-helix

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Fingerprint Dive into the research topics of 'A single amino acid substitution in the C terminus of OmpR alters DNA recognition and phosphorylation'. Together they form a unique fingerprint.

  • Cite this