Analysis of the Campylobacter jejuni FlgR response regulator suggests integration of diverse mechanisms to activate an NtrC-like protein

Stephanie N. Joslin, David R. Hendrixson

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

Flagellar motility in Campylobacter jejuni mediates optimal interactions with human or animal hosts. σ54 and the FlgSR two-component system are necessary for the expression of many C. jejuni flagellar genes. The FlgR response regulator is homologous to the NtrC family of transcriptional activators. These regulators usually contain an N-terminal receiver domain, a central domain that interacts with σ54 and hydrolyzes ATP, and a DNA-binding C-terminal domain. Most often, phosphorylation of the receiver domain influences its inherent ability to either positively or negatively control the activity of the regulator. In this study, we performed genetic and biochemical analyses to understand how FlgR activity is controlled to culminate in the expression of σ54-dependent flagellar genes. Our data suggest that the FlgR receiver domain has the capacity for both positive and negative regulation in controlling the activation of the protein. Analysis of the C-terminal domain of FlgR revealed that it lacks a DNA-binding motif and is not required for σ54-dependent flagellar gene expression. Further analysis of FlgR lacking the C-terminal domain indicates that this protein is partially functional in the absence of the cognate sensor kinase, FlgS, but its activity is still dependent on the phosphorylated residue in the receiver domain, D51. We hypothesize that the C-terminal domain may not function to bind DNA but may ensure the specificity of the phosphorylation of FlgR by FlgS. Our results demonstrate that FlgR activation mechanisms are unusual among characterized NtrC-like proteins and emphasize that various means are utilized by the NtrC family of proteins to control the transcription of target genes.

Original languageEnglish (US)
Pages (from-to)2422-2433
Number of pages12
JournalJournal of bacteriology
Volume190
Issue number7
DOIs
StatePublished - Apr 2008

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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