Single-stranded DNA binding protein and DNA helicase of bacteriophage T7 mediate homologous DNA strand exchange

EMBO J. 1996 Apr 15;15(8):2010-9.

Abstract

Two proteins encoded by bacteriophage T7, the gene 2.5 single-stranded DNA binding protein and the gene 4 helicase, mediate homologous DNA strand exchange. Gene 2.5 protein stimulates homologous base pairing of two DNA molecules containing complementary single-stranded regions. The formation of a joint molecule consisting of circular, single-stranded M13 DNA, annealed to homologous linear, duplex DNA having 3'- or 5'-single-stranded termini of approximately 100 nucleotides requires stoichiometric amounts of gene 2.5 protein. In the presence of gene 4 helicase, strand transfer proceeds at a rate of > 120 nucleotides/s in a polar 5' to 3' direction with respect to the invading strand, resulting in the production of circular duplex M13 DNA. Strand transfer is coupled to the hydrolysis of a nucleoside 5'-triphosphate. The reaction is dependent on specific interactions between gene 2.5 protein and gene 4 protein.

Publication types

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

MeSH terms

  • Bacteriophage T7 / genetics*
  • Bacteriophage T7 / metabolism*
  • DNA Helicases / genetics*
  • DNA Helicases / metabolism*
  • DNA Primase
  • DNA, Viral / genetics*
  • DNA, Viral / metabolism*
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / metabolism*
  • Models, Genetic
  • Protein Binding
  • RNA Nucleotidyltransferases / genetics*
  • RNA Nucleotidyltransferases / metabolism*
  • Recombination, Genetic*
  • Viral Proteins / genetics*
  • Viral Proteins / metabolism*

Substances

  • DNA, Viral
  • DNA-Binding Proteins
  • Viral Proteins
  • gene 2.5 protein, Enterobacteria phage T7
  • DNA Primase
  • RNA Nucleotidyltransferases
  • DNA Helicases