Regulated communication between the upstream face of RNA polymerase and the beta' subunit jaw domain

EMBO J. 2004 Oct 27;23(21):4264-74. doi: 10.1038/sj.emboj.7600407. Epub 2004 Oct 7.

Abstract

We used bacteriophage T7-encoded transcription inhibitor gene protein 2 (gp2) as a probe to study the contribution of the Escherichia coli RNA polymerase (RNAP) beta' subunit jaw domain--the site of gp2 binding--to activator and ATP hydrolysis-dependent open complex formation by the sigma(54)-RNAP. We show that, unlike sigma(70)-dependent transcription, activated transcription by sigma(54)-RNAP is resistant to gp2. In contrast, activator and ATP hydrolysis-independent transcription by sigma(54)-RNAP is highly sensitive to gp2. We provide evidence that an activator- and ATP hydrolysis-dependent conformational change involving the beta' jaw domain and promoter DNA is the basis for gp2-resistant transcription by sigma(54)-RNAP. Our results establish that accessory factors bound to the upstream face of the RNAP, communicate with the beta' jaw domain, and that such communication is subjected to regulation.

Publication types

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

MeSH terms

  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • DNA, Bacterial / metabolism
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • DNA-Directed RNA Polymerases / chemistry*
  • DNA-Directed RNA Polymerases / genetics
  • DNA-Directed RNA Polymerases / metabolism*
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism
  • Gene Expression Regulation, Bacterial
  • Macromolecular Substances
  • Models, Molecular
  • Nucleic Acid Denaturation
  • Promoter Regions, Genetic
  • Protein Binding
  • Protein Conformation
  • Protein Subunits / chemistry*
  • Protein Subunits / genetics
  • Protein Subunits / metabolism*
  • RNA Polymerase Sigma 54
  • Repressor Proteins / metabolism
  • Sigma Factor / genetics
  • Sigma Factor / metabolism
  • Transcription, Genetic

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • DNA-Binding Proteins
  • Escherichia coli Proteins
  • Macromolecular Substances
  • Protein Subunits
  • Repressor Proteins
  • Sigma Factor
  • gene 2 product, bacteriophage T7
  • rpoN protein, E coli
  • DNA-Directed RNA Polymerases
  • RNA Polymerase Sigma 54