Abstract
B. subtilis phage phi 29 has a terminal protein, p3, covalently linked to the 5' ends of the DNA through a phosphodiester bond between a serine residue and 5'-dAMP. This protein acts as a primer in DNA replication by forming an initiation complex with the 5'-terminal nucleotide dAMP. The amino acid sequence of the terminal protein, deduced from the nucleotide sequence of gene 3, showed the presence of 18 serine residues in a total of 266 amino acids. In this paper we have identified the serine involved in the linkage with the DNA as the residue 232, located close to the C-terminus of the molecule. This result was obtained by amino acid analysis of the peptide that remains linked to the DNA after proteinase K digestion of the terminal protein-phi 29 DNA complex and automated Edman degradation of the corresponding [125I]-labeled tryptic peptide. Prediction of the secondary structure of the terminal protein suggested that the serine residue involved in the linkage with the DNA is placed in a beta-turn, probably located on the external part of the molecule, as indicated by hydropathic values.
MeSH Terms
Amino Acid Sequence
Bacillus subtilis/genetics
Bacteriophages/genetics
Binding Sites
DNA, Viral/metabolism
DNA-Binding Proteins/metabolism
Iodine Radioisotopes
Peptide Fragments/analysis
Protein Binding
Serine
Trypsin
Viral Proteins/metabolism
Chemicals
DNA, Viral
DNA-Binding Proteins
Iodine Radioisotopes
Peptide Fragments
Viral Proteins
Serine
Trypsin
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Hermoso J M
Méndez E
Soriano F
Salas M
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