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Discriminatory function of ribonuclease H in the selective initiation of plasmid DNA replication.
Nucleic Acids Res. 1982 Feb 11; 10(3):833-53.NA

Abstract

The initiation stage of ColE1-type plasmid replication was reconstituted with purified protein fractions from Escherichia coli. The reconstituted system included DNA polymerase I, DNA ligase, RNA polymerase, DNA gyrase, and a discriminating activity copurifying with RNAase H (but free of RNAase III). Initiation of DNA synthesis in the absence of RNAase H did not occur at the normal replication origin and was non-selective with respect to the plasmid template. In the presence of RNAase H the system was selective for ColE1-type plasmids and could not accept the DNA of non-amplifiable plasmids. Electron microscopic analysis of the reaction product formed under discriminatory conditions indicated that origin usage and directionally of ColE1, RSF1030, and CloDF13 replication were consistent with the normal replication pattern of these plasmids. It is proposed that the initiation of ColE1-type replication depends on the formation of an extensive secondary structure in the origin primer RNA that prevents its degradation by RNAase H.

Authors

No affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

6278438

Citation

Hillenbrand, G, and W L. Staudenbauer. "Discriminatory Function of Ribonuclease H in the Selective Initiation of Plasmid DNA Replication." Nucleic Acids Research, vol. 10, no. 3, 1982, pp. 833-53.
Hillenbrand G, Staudenbauer WL. Discriminatory function of ribonuclease H in the selective initiation of plasmid DNA replication. Nucleic Acids Res. 1982;10(3):833-53.
Hillenbrand, G., & Staudenbauer, W. L. (1982). Discriminatory function of ribonuclease H in the selective initiation of plasmid DNA replication. Nucleic Acids Research, 10(3), 833-53.
Hillenbrand G, Staudenbauer WL. Discriminatory Function of Ribonuclease H in the Selective Initiation of Plasmid DNA Replication. Nucleic Acids Res. 1982 Feb 11;10(3):833-53. PubMed PMID: 6278438.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Discriminatory function of ribonuclease H in the selective initiation of plasmid DNA replication. AU - Hillenbrand,G, AU - Staudenbauer,W L, PY - 1982/2/11/pubmed PY - 1982/2/11/medline PY - 1982/2/11/entrez SP - 833 EP - 53 JF - Nucleic acids research JO - Nucleic Acids Res. VL - 10 IS - 3 N2 - The initiation stage of ColE1-type plasmid replication was reconstituted with purified protein fractions from Escherichia coli. The reconstituted system included DNA polymerase I, DNA ligase, RNA polymerase, DNA gyrase, and a discriminating activity copurifying with RNAase H (but free of RNAase III). Initiation of DNA synthesis in the absence of RNAase H did not occur at the normal replication origin and was non-selective with respect to the plasmid template. In the presence of RNAase H the system was selective for ColE1-type plasmids and could not accept the DNA of non-amplifiable plasmids. Electron microscopic analysis of the reaction product formed under discriminatory conditions indicated that origin usage and directionally of ColE1, RSF1030, and CloDF13 replication were consistent with the normal replication pattern of these plasmids. It is proposed that the initiation of ColE1-type replication depends on the formation of an extensive secondary structure in the origin primer RNA that prevents its degradation by RNAase H. SN - 0305-1048 UR - https://www.unboundmedicine.com/medline/citation/6278438/Discriminatory_function_of_ribonuclease_H_in_the_selective_initiation_of_plasmid_DNA_replication_ L2 - https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/10.3.833 DB - PRIME DP - Unbound Medicine ER -