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Closed Access item Characterization of a protein covalently linked to the 5′ termini of the DNA of Bacillus subtilis phage φ29

Authors:Salas, Margarita
Mellado, Rafael P.
Viñuela, Eladio
Sogo, José M.
Issue Date:25-Feb-1978
Citation:Journal of Biological Chemistry 119(2): 269-291 (1978)
Abstract:We have isolated a covalent DNA-protein complex from bacteriophage φ29 particles. Polyacrylamide gel electrophoresis and tryptic peptide analysis showed that the protein present in the complex is very similar or identical to p3, an early induced protein essential for viral DNA replication. When the DNA-protein complex is treated with the restriction endonuclease EcoRI, the proteinis specifically associated to the two terminal fragments, A and C. The protein is probably linked to the 5′ termini of the DNA since proteinase K-treated DNA is resistant to phosphorylation with polynucleotide kinase, even after treatment with alkaline phosphatase, while it is sensitive to exonuclease III. By electron microscopy the protein is visualized as a dot located at the ends of unit length DNA molecules. Mixed infection of Bacillus subtilis, at 42 °C, with ts† mutants in cistrons 2 and 3 only produces ts 2 progeny. This finding suggests that an inactive protein p3 bound to the DNA of the ts 3 mutant is not replaced by a functional protein and, as a consequence, replication of the ts 3 DNA does not occur.
Publisher version (URL):http://dx.doi.org/10.1016/0022-2836(78)90438-2
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