Phosphorylation alters the interaction of the response regulator OmpR with its sensor kinase EnvZ

Kirsten Mattison, Linda Kenney

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

OmpR and EnvZ comprise a two-component system that regulates the porin genes ompF and ompC in response to changes in osmolarity. EnvZ is autophosphorylated by intracellular ATP on a histidine residue, and it transfers the phosphoryl group to an aspartic acid residue of OmpR. EnvZ can also dephosphorylate phospho-OmpR (OmpR-P) to control the cellular level of OmpR-P. At low osmolarity, OmpR-P levels are low because of either low EnvZ kinase or high EnvZ phosphatase activities. At high osmolarity, OmpR-P is elevated. It has been proposed that EnvZ phosphatase is the activity that is regulated by osmolarity. OmpR is a two-domain response regulator; phosphorylation of OmpR increases its affinity for DNA, and DNA binding stimulates phosphorylation. The step that is affected by DNA depends upon the phosphodonor employed. In the present work, we have used fluorescence anisotropy and phosphotransfer assays to examine OmpR interactions with EnvZ. Our results indicate that phosphorylation greatly reduces the affinity of OmpR for the kinase, whereas DNA does not affect their interaction. The results presented cast serious doubts on the role of the EnvZ phosphatase in response to signaling in vivo.

Original languageEnglish (US)
Pages (from-to)11143-11148
Number of pages6
JournalJournal of Biological Chemistry
Volume277
Issue number13
DOIs
StatePublished - Mar 29 2002
Externally publishedYes

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Phosphorylation
Phosphoric Monoester Hydrolases
Osmolar Concentration
Phosphotransferases
DNA
Sensors
Polynucleotide 5'-Hydroxyl-Kinase
Porins
Histidine
Aspartic Acid
Fluorescence Polarization
Assays
Anisotropy
Genes
Adenosine Triphosphate
Fluorescence

ASJC Scopus subject areas

  • Biochemistry

Cite this

Phosphorylation alters the interaction of the response regulator OmpR with its sensor kinase EnvZ. / Mattison, Kirsten; Kenney, Linda.

In: Journal of Biological Chemistry, Vol. 277, No. 13, 29.03.2002, p. 11143-11148.

Research output: Contribution to journalArticle

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