Homologous pairing in vitro initiated by DNA synthesis

Abstract

A number of models have been proposed for the initiation of general genetic recombination. One of these, originally proposed by Meselson and Radding, imagines that the single-stranded 5′ tail that results from strand displacement DNA repair synthesis can initiate homologous recombination by invading a homologous duplex. The resultant D-loop intermediate is then processed into mature products. We demonstrate here that an in vitro system composed of the bacteriophage T4 uvsX protein (a RecA-like “strand transferase”) and part of the T4 DNA polymerase holoenzyme efficiently mediates pairing between nicked double-stranded circular and linear duplex DNAs, thereby demonstrating the feasibility of a key step in the Meselson-Radding model.