The characterization of a complex of three bacteriophage T4 recombination proteins, uvsX protein, uvsY protein, and gene 32 protein, on single-stranded DNA.

Abstract

Three recombination proteins of bacteriophage T4, uvsX, uvsY, and gene 32 proteins, were examined for the formation of a complex with short single-stranded DNA (ssDNA) molecules containing either 24 or 69 nucleotides. Gel-shift assays revealed that either the uvsX or uvsY protein, when present alone, formed a stable complex only with the 69-mer, while the gene 32 protein bound stably to both ssDNAs. However, a characteristic stable complex formed on the 24-mer when both the uvsX and uvsY proteins were present, and the uvsY protein bound to this DNA in the presence of the gene 32 protein. Isolation of the complexes by centrifugation through a glycerol gradient revealed their protein constituents and showed that the uvsX protein-uvsY protein-24-mer ssDNA complex formed even in the presence of excess gene 32 protein. The possible biological significance of these protein-DNA complexes is discussed.