Structural and mechanistic basis for the inhibition of Escherichia coli RNA polymerase by T7 Gp2

Mol Cell. 2012 Sep 14;47(5):755-66. doi: 10.1016/j.molcel.2012.06.013. Epub 2012 Jul 19.


The T7 phage-encoded small protein Gp2 is a non-DNA-binding transcription factor that interacts with the jaw domain of the Escherichia coli (Ec) RNA polymerase (RNAp) β' subunit and inhibits transcriptionally proficient promoter-complex (RPo) formation. Here, we describe the high-resolution solution structure of the Gp2-Ec β' jaw domain complex and show that Gp2 and DNA compete for binding to the β' jaw domain. We reveal that efficient inhibition of RPo formation by Gp2 requires the amino-terminal σ(70) domain region 1.1 (R1.1), and that Gp2 antagonizes the obligatory movement of R1.1 during RPo formation. We demonstrate that Gp2 inhibits RPo formation not just by steric occlusion of the RNAp-DNA interaction but also through long-range antagonistic effects on RNAp-promoter interactions around the RNAp active center that likely occur due to repositioning of R1.1 by Gp2. The inhibition of Ec RNAp by Gp2 thus defines a previously uncharacterized mechanism by which bacterial transcription is regulated by a viral factor.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Bacterial / chemistry
  • DNA, Bacterial / drug effects
  • DNA-Directed RNA Polymerases / antagonists & inhibitors*
  • DNA-Directed RNA Polymerases / chemistry
  • DNA-Directed RNA Polymerases / metabolism
  • Escherichia coli / enzymology*
  • Models, Molecular
  • Promoter Regions, Genetic / drug effects
  • Promoter Regions, Genetic / genetics
  • Protein Conformation
  • Repressor Proteins / chemistry
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism*
  • Transcription, Genetic / drug effects
  • Transcription, Genetic / genetics
  • Transcriptional Activation / drug effects
  • Transcriptional Activation / genetics


  • DNA, Bacterial
  • Repressor Proteins
  • gene 2 product, bacteriophage T7
  • DNA-Directed RNA Polymerases

Associated data

  • PDB/2LMC