Integration of multiple domains in a two-component sensor protein: the Bordetella pertussis BvgAS phosphorelay

EMBO J. 1996 Mar 1;15(5):1028-36.

Abstract

BvgS and BvgA, a two-component system, regulate virulence gene expression in Bordetella pertussis. BvgS is a transmembrane sensor protein that can autophosphorylate and phosphorylate BvgA. Phosphorylated BvgA activates transcription of virulence genes. The cytoplasmic region of BvgS contains three domains separated by alanine/proline-rich sequences--the transmitter, receiver and C-terminus. We report that the C-terminal domain, like the transmitter and receiver, is an essential part of the phosphorelay from BvgS to BvgA. The BvgS C-terminal domain is phosphorylated in trans via a phosphotransfer mechanism by the cytoplasmic portion of BvgS, and trans-phosphorylation of the C-terminal domain requires both the transmitter and receiver. We also demonstrate that phosphorylated, purified C-terminal domain alone is sufficient for phosphotransfer to BvgA. A point mutation in the C-terminal domain (His1172-->Gln) abolishes BvgS activity in vivo and eliminates detectable phosphorylation of BvgA in vitro. Activity of BvgS His 1172-->Gln could be restored by providing the wild-type C-terminal domain in trans. Our results indicate an obligatory role for an alternate phosphodonor module in the BvgAS phosphorelay.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Base Sequence
  • Bordetella pertussis / genetics
  • Bordetella pertussis / metabolism*
  • Bordetella pertussis / pathogenicity
  • DNA Primers / genetics
  • DNA, Bacterial / genetics
  • Escherichia coli / genetics
  • Escherichia coli Proteins*
  • Gene Expression Regulation, Bacterial
  • Genetic Complementation Test
  • Molecular Sequence Data
  • Molecular Structure
  • Mutagenesis, Site-Directed
  • Phosphorylation
  • Signal Transduction / genetics
  • Signal Transduction / physiology
  • Transcription Factors / chemistry
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Virulence / genetics
  • Virulence / physiology

Substances

  • Bacterial Proteins
  • BvgA protein, Bacteria
  • DNA Primers
  • DNA, Bacterial
  • Escherichia coli Proteins
  • EvgA protein, E coli
  • Transcription Factors
  • bvgS protein, Bordetella pertussis