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PMID: 6278438 Published · ppublish English Comparative Study Journal Article

Discriminatory function of ribonuclease H in the selective initiation of plasmid DNA replication.

Nucleic acids research ·Vol. 10 ·No. 3 ·1982-02-11 ·Pages 833-53

Hillenbrand G, Staudenbauer WL

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.

MeSH Terms
DNA Polymerase I/metabolism DNA Replication DNA, Recombinant/metabolism Endonucleases/metabolism Escherichia coli/genetics,metabolism Microscopy, Electron Mutation Plasmids Ribonuclease H Ribonucleases/metabolism Species Specificity Templates, Genetic
Chemicals
DNA, Recombinant DNA Polymerase I Endonucleases Ribonucleases Ribonuclease H
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Hillenbrand G
Staudenbauer W L
References (41)
41 references, click to expand
  1. Description of an incompatibility mutant of Escherichia coli.
    J Bacteriol. 1973 Jul;115(1):213-20 PMID: 4577742
  2. General method for the isolation of plasmid deoxyribonucleic acid.
    J Bacteriol. 1973 Nov;116(2):1064-6 PMID: 4583233
  3. Replication of colicin E1 plasmid DNA in minicells from a unique replication initiation site.
    Proc Natl Acad Sci U S A. 1974 Jun;71(6):2256-9 PMID: 4601583
  4. Unidirectional replication of plasmid ColE1 DNA.
    Nature. 1974 Sep 27;251(5473):337-40 PMID: 4610399
  5. The degree of unwinding of the DNA helix by ethidium. I. Titration of twisted PM2 DNA molecules in alkaline cesium chloride density gradients.
    J Mol Biol. 1974 Nov 15;89(4):783-801 PMID: 4449133
  6. Priming of superhelical SV40 DNA by Escherichia coli RNA polymerase for in vitro DNA synthesis.
    Biochemistry. 1975 Jan 28;14(2):307-16 PMID: 164202
  7. A simple method for the preparation of large quantities of pure plasmid DNA.
    Biochim Biophys Acta. 1975 Apr 2;383(4):457-63 PMID: 1092355
  8. Cloning, isolation, and characterization of replication regions of complex plasmid genomes.
    Proc Natl Acad Sci U S A. 1975 Jun;72(6):2242-6 PMID: 1094460
  9. Ten proteins required for conversion of phiX174 single-stranded DNA to duplex form in vitro. Resolution and reconstitution.
    J Biol Chem. 1975 Aug 10;250(15):5859-65 PMID: 1097445
  10. dnaG gene product, a rifampicin-resistant RNA polymerase, initiates the conversion of a single-stranded coliphage DNA to its duplex replicative form.
    J Biol Chem. 1975 Aug 10;250(15):5995-6001 PMID: 1097446
  11. phiX174 DNA-dependent DNA synthesis in vitro: requirement for P1 ban protein in dnaB mutant extracts of Escherichia coli.
    Proc Natl Acad Sci U S A. 1975 Oct;72(10):3907-11 PMID: 670
  12. Reconstruction of bacteriophage T4 DNA replication apparatus from purified components: rolling circle replication following de novo chain initiation on a single-stranded circular DNA template.
    Proc Natl Acad Sci U S A. 1975 Dec;72(12):4800-4 PMID: 1061070
  13. DNA gyrase: an enzyme that introduces superhelical turns into DNA.
    Proc Natl Acad Sci U S A. 1976 Nov;73(11):3872-6 PMID: 186775
  14. Selective inhibition of in vitro DNA synthesis dependent on phiX174 compared with fd DNA. I. Protein requirements for selective inhibition.
    J Biol Chem. 1977 Apr 25;252(8):2524-33 PMID: 140166
  15. Selective inhibition of phiX RFII compared with fd RFII DNA synthesis in vitro. II. Resolution of discrimination reaction into multiple steps.
    J Biol Chem. 1977 Apr 25;252(8):2534-44 PMID: 323248
  16. Origin of replication of colicin E1 plasmid DNA.
    Proc Natl Acad Sci U S A. 1977 May;74(5):1865-9 PMID: 325558
  17. The nucleotide sequence surrounding the origin of DNA replication of Col E1.
    Nucleic Acids Res. 1977 Sep;4(9):3123-42 PMID: 410007
  18. Studies on the transcription complex of Escherichia coli RNA polymerase.
    Eur J Biochem. 1977 Oct 3;79(2):401-9 PMID: 336358
  19. Replication of the ampicillin resistance plasmid RSF1030 in extracts of Escherichia coli: separation of the replication cycle into early and late stages.
    Mol Gen Genet. 1977 Nov 4;156(1):27-34 PMID: 340889
  20. Origin of replication of pBR345 plasmid DNA.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5265-9 PMID: 271952
  21. Refinement of the coomassie blue method of protein quantitation. A simple and linear spectrophotometric assay for less than or equal to 0.5 to 50 microgram of protein.
    Anal Biochem. 1978 May;86(1):142-6 PMID: 655375
  22. Structure and replication of the colicin E1 plasmid.
    Curr Top Microbiol Immunol. 1978;83:93-156 PMID: 365473
  23. Origin and direction of replication of the bacteriocinogenic plasmid Clo DF13.
    Nucleic Acids Res. 1979 Jan;6(1):71-80 PMID: 370788
  24. Origin and direction of DNA replication of plasmid RSF1030.
    Proc Natl Acad Sci U S A. 1979 Feb;76(2):736-40 PMID: 370835
  25. Initiation of DNA synthesis in Escherichia coli.
    Annu Rev Biochem. 1979;48:999-1034 PMID: 382998
  26. Initiation of replication of plasmid ColE1 DNA by RNA polymerase, ribonuclease H, and DNA polymerase I.
    Cold Spring Harb Symp Quant Biol. 1979;43 Pt 1:409-17 PMID: 225109
  27. Genetic and physical studies on the replication of ColE1-type plasmids.
    Cold Spring Harb Symp Quant Biol. 1979;43 Pt 1:69-76 PMID: 383386
  28. Complete nucleotide sequence of the Escherichia coli plasmid pBR322.
    Cold Spring Harb Symp Quant Biol. 1979;43 Pt 1:77-90 PMID: 383387
  29. Essential regions for replication of a stringent and a relaxed plasmid in Escherichia coli.
    Cold Spring Harb Symp Quant Biol. 1979;43 Pt 1:99-103 PMID: 383389
  30. Characterization of an improved in vitro DNA replication system for Escherichia coli plasmids.
    Nucleic Acids Res. 1979 Jul 25;6(10):3289-304 PMID: 384367
  31. Role of plasmid-coded RNA and ribonuclease III in plasmid DNA replication.
    Cell. 1979 Sep;18(1):61-71 PMID: 389434
  32. Regulation of initiation of DNA replication.
    Annu Rev Genet. 1979;13:355-91 PMID: 395895
  33. Formation of an RNA primer for initiation of replication of ColE1 DNA by ribonuclease H.
    Proc Natl Acad Sci U S A. 1980 May;77(5):2450-4 PMID: 6156450
  34. The nucleotide sequence surrounding the replication origin of the cop3 mutant of the bacteriocinogenic plasmid Clo DF13.
    Nucleic Acids Res. 1980 Apr 11;8(7):1459-73 PMID: 6253936
  35. Plasmid R1 DNA replication dependent on protein synthesis in cell-free extracts of E. coli.
    Nature. 1981 Jan 22;289(5795):326-8 PMID: 7005697
  36. Regulation of plasmid DNA synthesis: isolation and characterization of copy number mutants of miniR6-5 and miniF plasmids.
    Mol Gen Genet. 1981;181(1):29-35 PMID: 6261084
  37. An Escherichia coli mutant thermosensitive in the B subunit of DNA gyrase: effect on the structure and replication of the colicin E1 plasmid in vitro.
    Mol Gen Genet. 1981;181(1):52-6 PMID: 6261085
  38. Replication and structure of the bacteriocinogenic plasmids Clo DF13 and CoI E1.
    Plasmid. 1981 Jan;5(1):76-99 PMID: 6164067
  39. DNA gyrase: affinity chromatography on novobiocin-Sepharose and catalytic properties.
    Nucleic Acids Res. 1981 Aug 11;9(15):3589-603 PMID: 6269086
  40. Site-specific interaction of DNA gyrase with DNA.
    Proc Natl Acad Sci U S A. 1981 Jul;78(7):4165-9 PMID: 6270661
  41. The nucleotide sequence of the replication origin of plasmid NTP1.
    Nucleic Acids Res. 1981 Sep 11;9(17):4355-66 PMID: 6272204
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/10.3.833
Published
1982-02-11
Pages
833-53
Language
English
Region
England
NLM ID
0411011
PMCID
PMC326206
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/10.3.833 PMC Full Text View journal details
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