A comparison of the DNA binding and bending capacities and the oligomeric states of the immunity repressors of heteroimmune coliphages P2 and WPhi

Nucleic Acids Res. 2007;35(10):3167-80. doi: 10.1093/nar/gkm171. Epub 2007 May 7.

Abstract

Bacteriophages P2 and WPhi are heteroimmune members of the P2-like family of temperate Escherichia coli phages. Temperate phages can grow lytically or form lysogeny after infection. A transcriptional switch that contains two con-vergent promoters, Pe and Pc, and two repressors regulate what life mode to enter. The immunity repressor C is the first gene of the lysogenic operon, and it blocks the early Pe promoter. In this work, some characteristics of the C proteins of P2 and WPhi are compared. An in vivo genetic analysis shows that WPhi C, like P2 C, has a strong dimerization activity in the absence of its DNA target. Both C proteins recognize two directly repeated sequences, termed half-sites and a strong bending is induced in the respective DNA target upon binding. P2 C is unable to bind to one half-site as opposed to WPhi, but both half-sites are required for repression of WPhi Pe. A reduction from three to two helical turns between the centers of the half-sites in WPhi has no significant effect on the capacity to repress Pe. However, the protein-DNA complexes formed differ, as determined by electrophoretic mobility shift experiments. A difference in spontaneous phage production is observed in isogenic lysogens.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacteriophage P2 / genetics*
  • Bacteriophage P2 / growth & development
  • Binding Sites
  • Coliphages / genetics*
  • Coliphages / growth & development
  • DNA / chemistry
  • DNA / metabolism
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / metabolism
  • Dimerization
  • Gene Expression Regulation, Viral
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Operator Regions, Genetic*
  • Promoter Regions, Genetic
  • Repressor Proteins / chemistry
  • Repressor Proteins / metabolism*
  • Viral Proteins / chemistry
  • Viral Proteins / metabolism*

Substances

  • C protein, Enterobacteria phage P2
  • DNA-Binding Proteins
  • Repressor Proteins
  • Viral Proteins
  • DNA