Identification of the promoter region and the transcriptional regulatory sequence of the evgAS operon of Escherichia coli

Biosci Biotechnol Biochem. 1998 Feb;62(2):286-90. doi: 10.1271/bbb.62.286.

Abstract

The evgAS operon of Escherichia coli encodes the EvgA response regulator and the EvgS sensory kinase, which are members of one of the two-component signal transduction systems of Escherichia coli. In this study, we identified the evg promoter and the EvgA-responsive element. Primer extension analysis found two evg transcriptional initiation sites, designated P1 (+1) and P2 (-10), and placed them 114 bp and 124 bp upstream of evgA, respectively. A gel retardation assay demonstrated that EvgA specifically bound to an inverted repeat located between -102 and -128 counting from P1. We also did a beta-galactosidase induction experiment using a promoter-probing vector and found that the EvgA-binding sequence was important to stimulate the evg promoter. These results suggest that the expression of evgAS is positively regulated by its own product, EvgA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Base Sequence
  • DNA Primers / chemistry
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / metabolism
  • Electrophoresis, Agar Gel
  • Escherichia coli / chemistry
  • Escherichia coli / genetics*
  • Escherichia coli Proteins*
  • Gene Expression Regulation, Bacterial*
  • Molecular Sequence Data
  • Operon / genetics*
  • Polymerase Chain Reaction
  • Promoter Regions, Genetic*
  • Recombinant Fusion Proteins / chemistry
  • Restriction Mapping
  • Signal Transduction / genetics
  • Transcription Factors / chemistry
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Transcription, Genetic
  • beta-Galactosidase / analysis

Substances

  • Bacterial Proteins
  • BvgA protein, Bacteria
  • DNA Primers
  • DNA, Bacterial
  • Escherichia coli Proteins
  • EvgA protein, E coli
  • Recombinant Fusion Proteins
  • Transcription Factors
  • bvgS protein, Bordetella pertussis
  • beta-Galactosidase