Location of the Serine Residue Involved in the Linkage Between the Terminal Protein and the DNA of Phage Phi 29

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1985 Nov 11

PMID 3934646

Citations 38

Authors

J M Hermoso

E Mendez

F Soriano

M Salas

Affiliations

Soon will be listed here.

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.

Citing Articles

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DNA-Binding Proteins Essential for Protein-Primed Bacteriophage Φ29 DNA Replication.

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Disclosing early steps of protein-primed genome replication of the Gram-positive tectivirus Bam35.

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Dissecting the role of the ϕ29 terminal protein DNA binding residues in viral DNA replication.

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Mutational analysis of the terminal protein Tpg of Streptomyces chromosomes: identification of the deoxynucleotidylation site.

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