Strand Exchange Through a DNA-Protein Complex Requires a DNA Helicase

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.