Abstract
Gene 59 of bacteriophage T4 is involved in genetic recombination and recombination-dependent DNA replication. We purified the protein encoded by gene 59 after cloning the gene in an expression vector. The molecular weight and the sequence of seven N-terminal amino acids of the protein were consistent with the nucleotide sequence of the gene (Hahn, S., Kruse, U., and Ruger, W. (1986) Nucleic Acids Res. 14, 9311-9327). The purified 59 protein appeared to be a monomer under nondenaturing conditions, and it bound to single-stranded DNA in preference to double-stranded DNA. The protein could bind to a 24-nucleotide-long single-stranded DNA molecule that had previously bound either 32 protein (single-stranded DNA-binding protein) or uvsY protein (enhancer of uvsX protein), but it would not bind to this DNA if it had previously bound uvsX protein (synaptase). The binding occurred rapidly with 32 protein-bound DNA and slowly with uvsY protein-bound DNA. Accordingly, 32 protein bound specifically to an agarose matrix containing immobilized 59 protein. The 41 protein (DNA helicase) also bound specifically to this agarose matrix.
Highlights
Gene 69 of bacteriophage T4 is involved in genetic and Minagawa, 1989) as well as an essential role in DNA recombinatioannrdecombination-dependent
We purified the protein encoded by gene Gene 59 is another gene thought to promote DNA recom
Purification of the 59 Protein-As expected from the amino acid sequence deduced from nucleotide sequence (Hahn aetl., 1986),E. coli cells harboring pUC19-59 accumulated a 26-kDa protein detectable by SDS-PAGE andCoomassie Blue staining
Summary
From the Departmentof Biology, College of General Education, Osaka University, Osaka560, Japan. We purified the protein encoded by gene Gene 59 is another gene thought to promote DNA recom-. Recombination and replication are, bound to anagarosematrixcontaining intimately interrelated and depend on each other in the T4 immobilized 69 protein. The DNA recombination of bacteriophage T4 is catalyzed independently purified the protein; they have shown that the by its own gene products. The genes include UUSXu,vsY, 32, protein rescues the DNA-dependent GTP hydrolysis activity. UvsX protein binds cooperatively to ssDNA' and of 41 protein (DNA helicase) from inhibition by protein catalyzes transfer of the ssDNA intoa homologousDNA and that,by loading 41 protein onto ssDNA, it enhances the duplex 1986).UvsY protein promotes the binding of uvsX protein to data is consistent with their results and helps to explain the ssDNA that is covered with the ssDNA-binding protein of molecular basis of 59 protein function
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