DOI: 10.1073/pnas.79.18.5522. Initiation of phage phi 29 DNA replication in vitro: formation of a covalent complex between the terminal protein, p3, and 5'-dAMP.

Initiation of phage phi 29 DNA replication in vitro: formation of a covalent complex between the terminal protein, p3, and 5'-dAMP.

10.1073/pnas.79.18.5522

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Incubation of extracts of phi 29-infected Bacillus subtilis with [alpha-32P]dATP produced a labeled protein having the electrophoretic mobility of p3, the 5'-terminal protein of phi 29 DNA. The reaction product was resistant to treatment with micrococcal nuclease, phosphatase, and RNases A and T1 and sensitive to proteinase K. Incubation of the 32P-labeled protein with piperidine under conditions in which the phi 29 DNA-protein p3 linkage is hydrolyzed released 5'-dAMP. The reaction with [alpha-32P]dATP was strongly inhibited by anti-p3 serum and required the preence of phi 29 DNA-protein p3 complex; no reaction took place with proteinase K-treated phi29 DNA. These results, together with those of acid hydrolysis and partial proteolysis, indicated that a covalent complex between protein p3 and 5'-dAMP is formed in vitro. The initiation complex (protein p3-dAMP) formed in the presence of 0.5 microM [alpha-32P]dATP can be elongated by addition of 40 microM dNTPs. Treatment with piperidine of the product elongated in the presence of 2',3'-dideoxycytidine 5'-triphosphate released the expected oligonucleotides, 9 and 12 bases long, taking into account the sequence at the left and right DNA ends, respectively.

Bibliography

PeÃ±alva, M. A., & Salas, M. (1982). Initiation of phage phi 29 DNA replication in vitro: formation of a covalent complex between the terminal protein, p3, and 5â-dAMP. Proceedings of the National Academy of Sciences, 79(18), 5522â5526.

Authors 2

M A Peñalva (first)
M Salas (additional)

References 0 Referenced 96

None

Dates

Type	When
Created	19 years, 3 months ago (May 31, 2006, 4:53 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 11:18 a.m.)
Indexed	3 months, 2 weeks ago (May 18, 2025, 10:24 a.m.)
Issued	43 years ago (Sept. 1, 1982)
Published	43 years ago (Sept. 1, 1982)
Published Online	43 years ago (Sept. 1, 1982)
Published Print	43 years ago (Sept. 1, 1982)

Funders 0

None

BibTeX

@article{Pe_alva_1982, title={Initiation of phage phi 29 DNA replication in vitro: formation of a covalent complex between the terminal protein, p3, and 5’-dAMP.}, volume={79}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.79.18.5522}, DOI={10.1073/pnas.79.18.5522}, number={18}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Peñalva, M A and Salas, M}, year={1982}, month=sep, pages={5522–5526} }

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