Functional interactions between phage T4 and E. coli DNA-binding proteins during the presynapsis phase of homologous recombination.

The "protein machine" for phage T4 homologous recombination has begun to be assembled in vitro. A particularly heavily studied reaction has been the uvsX protein (a RecA-like strand transferase)-mediated homologous pairing reaction between single and double-stranded DNAs, a key step in the recombination cycle in vivo. A necessary prerequisite for uvsX protein-mediated pairing is the polymerization of this factor along the invading single strand, a process known as presynapsis. Recent work has indicated that at least two other T4 recombination factors are involved in this process as well, the uvsY and gene 32 products. These proteins are also ssDNA-binding factors and exhibit an affinity for UvsX and each other. In order to begin to sort out the potential functional roles played by these protein-protein interactions in presynapsis, I have examined the ability of the uvsX protein to form stable filaments along ssDNA in the presence of these proteins. It is shown that the uvsY protein relieves the inhibition to filament formation due to the presence of the gene 32 protein, but experiments with the E. coli SSB protein (the bacterial analogue of gp32) suggest that this effect does not involve a direct interaction between UvsY and gp32.