The response regulator OmpR oligomerizes via β-sheets to form head-to-head dimers

Ann E. Maris, Don Walthers, Kirsten Mattison, Nicole Byers, Linda Kenney

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

In Escherichia coli, the EnvZ/OmpR two-component regulatory system regulates expression of the porin genes ompF and ompC in response to changes in osmolarity. It has recently become apparent that OmpR functions as a global regulator, by regulating the expression of many genes in addition to the porin genes. OmpR consists of two domains; phosphorylation of the N-terminal receiver domain increases DNA binding affinity of the C-terminal domain and vice versa. Many response regulators including PhoB and FixJ dimerize upon phosphorylation. Here, we demonstrate that OmpR dimerization is stimulated by phosphorylation or by DNA binding. The dimerization interface revealed here was unanticipated and had previously not been predicted. Using the accepted head-to-tail tandem-binding model as a guide, we set out to examine the intermolecular interactions between OmpR dimers bound to DNA by protein-protein cross-linking methods. Surprisingly, amino acid positions that we expected to form cross-linked dimers did not. Conversely, positions predicted not to form dimers did. Because of these results, we designed a series of 23 cysteine-substituted OmpR mutants that were used to investigate dimer interfaces formed via the β-sheet region. This four-stranded β-sheet is a unique feature of the OmpR group of winged helix-turn-helix proteins. Many of the cysteine-substituted mutants are dominant to wild-type OmpR, are phosphorylated by acetyl phosphate as well as the cognate kinase EnvZ, and the cross-linked proteins are capable of binding to DNA. Our results are consistent with a model in which OmpR binds to DNA in a head-to-head orientation, in contrast to the previously proposed asymmetric head-to-tail model. They also raise the possibility that OmpR may be capable of adopting more than one orientation as it binds to a vast array of genes to activate or repress transcription.

Original languageEnglish (US)
Pages (from-to)843-856
Number of pages14
JournalJournal of Molecular Biology
Volume350
Issue number5
DOIs
StatePublished - Jul 29 2005
Externally publishedYes

Fingerprint

DNA
Porins
Phosphorylation
Dimerization
Cysteine
Gene Expression
Proteins
Protein Binding
Osmolar Concentration
Genes
Phosphotransferases
Escherichia coli
Amino Acids
acetyl phosphate

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology

Cite this

The response regulator OmpR oligomerizes via β-sheets to form head-to-head dimers. / Maris, Ann E.; Walthers, Don; Mattison, Kirsten; Byers, Nicole; Kenney, Linda.

In: Journal of Molecular Biology, Vol. 350, No. 5, 29.07.2005, p. 843-856.

Research output: Contribution to journalArticle

Maris, Ann E. ; Walthers, Don ; Mattison, Kirsten ; Byers, Nicole ; Kenney, Linda. / The response regulator OmpR oligomerizes via β-sheets to form head-to-head dimers. In: Journal of Molecular Biology. 2005 ; Vol. 350, No. 5. pp. 843-856.
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