The Sep1 strand exchange protein from Saccharomyces cerevisiae promotes a paranemic joint between homologous DNA molecules

Abstract

Strand exchange protein 1 (Sep1) from the yeast Saccharomyces cerevisiae promotes the transfer of one strand of a linear duplex DNA to a homologous single-stranded DNA circle. Using a nitrocellulose filter binding assay and electron microscopy, we find that Sep1 promotes the pairing of homologous DNA molecules via a paranemic joint. In this joint there is no net intertwining of the parental DNA molecules, as in the standard plectonemic double helix. The paranemic joints form with as little as 41 bp of homology between the parental DNA molecules. The substrates used were a circular molecule (either single-stranded DNA or duplex supercoiled DNA) and a linear duplex with heterologous regions at both ends to bar duplex plectonemic intertwining. We excluded the possibility that the exonuclease activity of Sep1 exposes complementary single-stranded regions that constitute the joint. The paranemic joint is the key intermediate in the search for homologous DNA by the RecA protein of Escherichia coli. Our results imply that the search process in a eukaryote such as yeast can be mechanistically similar.