Strand exchange through a DNA-protein complex requires a DNA helicase

Biochem Biophys Res Commun. 1994 Dec 15;205(2):1004-9. doi: 10.1006/bbrc.1994.2766.

Abstract

The phage T4 uvsX and gene 32 proteins are capable of mediating homologous strand exchange, a central reaction in general genetic recombination, in vitro using naked DNA substrates. However, strand exchange is blocked by a sequence specific DNA-protein complex. Since protein-complexed substrates must be employed in vivo, this suggests that another factor(s) is required for strand exchange with protein-complexed DNAs. We show here that a DNA helicase, the T4 dda protein, allows the phage recombination machinery to drive branch migration through a RNA polymerase-promoter complex. This is the first observation of in vitro strand exchange using protein-bound substrates. These results suggest that a DNA helicase is a necessary component of the "protein machine" that mediates recombination in vivo.

Publication types

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

MeSH terms

  • Adenosine Triphosphatases / metabolism*
  • Bacteriophage T4 / genetics
  • Bacteriophage T4 / metabolism
  • DNA / metabolism*
  • DNA Helicases / metabolism*
  • DNA-Binding Proteins / metabolism*
  • DNA-Directed RNA Polymerases / metabolism
  • Genes, Viral
  • Membrane Proteins / metabolism*
  • Promoter Regions, Genetic*
  • Protein Binding
  • Viral Proteins / metabolism*

Substances

  • DNA-Binding Proteins
  • Membrane Proteins
  • UvsX protein, Enterobacteria phage T4
  • Viral Proteins
  • gp32 protein, Enterobacteria phage T4
  • DNA
  • DNA-Directed RNA Polymerases
  • Adenosine Triphosphatases
  • DNA Helicases
  • dda DNA helicase protein, Bacteriophage T4