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Multiple mechanisms for initiation of ColE1 DNA replication: DNA synthesis in the presence and absence of ribonuclease H.
Cell. 1987 Dec 24; 51(6):1113-22.Cell

Abstract

A transcript (RNA II) of plasmid ColE1 that hybridizes with the template DNA is cleaved by RNAase H and used as a primer by DNA polymerase I. However, the plasmid can replicate in bacteria lacking both enzymes, apparently using a different mechanism of initiation of replication. Here we report in vivo and in vitro studies on initiation of DNA replication in the presence or absence of either or both enzymes. Hybridization of RNA II with the template DNA is always required for initiation. Hybridized RNA II is cleaved by RNAase H to form a primer or used as a primer without cleavage by RNAase H. Hybridization also creates a single-stranded region on the nontranscribed strand that can serve as a template for synthesis of the lagging strand in a reaction that does not require DNA polymerase I. Lagging strand synthesis terminates 17 nucleotides upstream of the normal replication origin, forcing unidirectional replication.

Authors+Show Affiliations

Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

2446774

Citation

Dasgupta, S, et al. "Multiple Mechanisms for Initiation of ColE1 DNA Replication: DNA Synthesis in the Presence and Absence of Ribonuclease H." Cell, vol. 51, no. 6, 1987, pp. 1113-22.
Dasgupta S, Masukata H, Tomizawa J. Multiple mechanisms for initiation of ColE1 DNA replication: DNA synthesis in the presence and absence of ribonuclease H. Cell. 1987;51(6):1113-22.
Dasgupta, S., Masukata, H., & Tomizawa, J. (1987). Multiple mechanisms for initiation of ColE1 DNA replication: DNA synthesis in the presence and absence of ribonuclease H. Cell, 51(6), 1113-22.
Dasgupta S, Masukata H, Tomizawa J. Multiple Mechanisms for Initiation of ColE1 DNA Replication: DNA Synthesis in the Presence and Absence of Ribonuclease H. Cell. 1987 Dec 24;51(6):1113-22. PubMed PMID: 2446774.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Multiple mechanisms for initiation of ColE1 DNA replication: DNA synthesis in the presence and absence of ribonuclease H. AU - Dasgupta,S, AU - Masukata,H, AU - Tomizawa,J, PY - 1987/12/24/pubmed PY - 1987/12/24/medline PY - 1987/12/24/entrez SP - 1113 EP - 22 JF - Cell JO - Cell VL - 51 IS - 6 N2 - A transcript (RNA II) of plasmid ColE1 that hybridizes with the template DNA is cleaved by RNAase H and used as a primer by DNA polymerase I. However, the plasmid can replicate in bacteria lacking both enzymes, apparently using a different mechanism of initiation of replication. Here we report in vivo and in vitro studies on initiation of DNA replication in the presence or absence of either or both enzymes. Hybridization of RNA II with the template DNA is always required for initiation. Hybridized RNA II is cleaved by RNAase H to form a primer or used as a primer without cleavage by RNAase H. Hybridization also creates a single-stranded region on the nontranscribed strand that can serve as a template for synthesis of the lagging strand in a reaction that does not require DNA polymerase I. Lagging strand synthesis terminates 17 nucleotides upstream of the normal replication origin, forcing unidirectional replication. SN - 0092-8674 UR - https://www.unboundmedicine.com/medline/citation/2446774/Multiple_mechanisms_for_initiation_of_ColE1_DNA_replication:_DNA_synthesis_in_the_presence_and_absence_of_ribonuclease_H_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0092-8674(87)90597-6 DB - PRIME DP - Unbound Medicine ER -