T7 phage factor required for managing RpoS in Escherichia coli

Proc Natl Acad Sci U S A. 2018 Jun 5;115(23):E5353-E5362. doi: 10.1073/pnas.1800429115. Epub 2018 May 22.

Abstract

T7 development in Escherichia coli requires the inhibition of the housekeeping form of the bacterial RNA polymerase (RNAP), Eσ70, by two T7 proteins: Gp2 and Gp5.7. Although the biological role of Gp2 is well understood, that of Gp5.7 remains to be fully deciphered. Here, we present results from functional and structural analyses to reveal that Gp5.7 primarily serves to inhibit EσS, the predominant form of the RNAP in the stationary phase of growth, which accumulates in exponentially growing E. coli as a consequence of the buildup of guanosine pentaphosphate [(p)ppGpp] during T7 development. We further demonstrate a requirement of Gp5.7 for T7 development in E. coli cells in the stationary phase of growth. Our finding represents a paradigm for how some lytic phages have evolved distinct mechanisms to inhibit the bacterial transcription machinery to facilitate phage development in bacteria in the exponential and stationary phases of growth.

Keywords: Escherichia coli; RNA polymerase; T7 phage; stationary phase; transcription regulation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / metabolism*
  • Bacteriophage T7 / enzymology
  • Bacteriophage T7 / genetics
  • Bacteriophage T7 / metabolism*
  • Crystallography, X-Ray
  • DNA-Directed DNA Polymerase / metabolism
  • DNA-Directed RNA Polymerases / antagonists & inhibitors*
  • DNA-Directed RNA Polymerases / metabolism
  • Escherichia coli / metabolism
  • Escherichia coli / virology*
  • Models, Molecular
  • Promoter Regions, Genetic
  • Protein Conformation
  • Repressor Proteins / metabolism*
  • Sigma Factor / metabolism*
  • Transcription, Genetic

Substances

  • Bacterial Proteins
  • Repressor Proteins
  • Sigma Factor
  • gene 2 product, bacteriophage T7
  • sigma factor KatF protein, Bacteria
  • bacteriophage T7 induced DNA polymerase
  • DNA-Directed RNA Polymerases
  • DNA-Directed DNA Polymerase

Associated data

  • PDB/6FI7