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. 1986 Mar;5(3):631–637. doi: 10.1002/j.1460-2075.1986.tb04257.x

Are single-stranded circles intermediates in plasmid DNA replication?

H te Riele, B Michel, S D Ehrlich
PMCID: PMC1166809  PMID: 3011418

Abstract

Plasmid pC194 exists as circular double-stranded and single-stranded DNA in Bacillus subtilis and Staphylococcus aureus. We report here that the plasmid pHV33, composed of pBR322 and pC194, exists as double- and single-stranded DNA in Escherichia coli, provided that the replication functions of pC194 are intact. Single-stranded pHV33 DNA is converted to double-stranded DNA by complementary strand synthesis probably initiated at rriB, a primosome assembly site present on pBR322. The efficiency of complementary strand synthesis affects the double-stranded copy number, which suggests that single-stranded DNA is a plasmid replication intermediate.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arai K., Kornberg A. Unique primed start of phage phi X174 DNA replication and mobility of the primosome in a direction opposite chain synthesis. Proc Natl Acad Sci U S A. 1981 Jan;78(1):69–73. doi: 10.1073/pnas.78.1.69. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arai K., Low R. L., Kornberg A. Movement and site selection for priming by the primosome in phage phi X174 DNA replication. Proc Natl Acad Sci U S A. 1981 Feb;78(2):707–711. doi: 10.1073/pnas.78.2.707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boeke J. D., Vovis G. F., Zinder N. D. Insertion mutant of bacteriophage f1 sensitive to EcoRI. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2699–2702. doi: 10.1073/pnas.76.6.2699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  5. Cashman J. S., Webster R. E., Steege D. A. Transcription of bacteriophage fl. The major in vivo RNAs. J Biol Chem. 1980 Mar 25;255(6):2554–2562. [PubMed] [Google Scholar]
  6. Dagert M., Jones I., Goze A., Romac S., Niaudet B., Ehrlich S. D. Replication functions of pC194 are necessary for efficient plasmid transduction by M13 phage. EMBO J. 1984 Jan;3(1):81–86. doi: 10.1002/j.1460-2075.1984.tb01764.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ehrlich S. D. DNA cloning in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1433–1436. doi: 10.1073/pnas.75.3.1433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Geider K., Beck E., Schaller H. An RNA transcribed from DNA at the origin of phage fd single strand to replicative form conversion. Proc Natl Acad Sci U S A. 1978 Feb;75(2):645–649. doi: 10.1073/pnas.75.2.645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Goze A., Ehrlich S. D. Replication of plasmids from Staphylococcus aureus in Escherichia coli. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7333–7337. doi: 10.1073/pnas.77.12.7333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gray C. P., Sommer R., Polke C., Beck E., Schaller H. Structure of the orgin of DNA replication of bacteriophage fd. Proc Natl Acad Sci U S A. 1978 Jan;75(1):50–53. doi: 10.1073/pnas.75.1.50. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Horinouchi S., Weisblum B. Nucleotide sequence and functional map of pC194, a plasmid that specifies inducible chloramphenicol resistance. J Bacteriol. 1982 May;150(2):815–825. doi: 10.1128/jb.150.2.815-825.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Horiuchi K., Ravetch J. V., Zinder N. D. DNA replication of bacteriophage f1 in vivo. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):389–399. doi: 10.1101/sqb.1979.043.01.045. [DOI] [PubMed] [Google Scholar]
  13. Koepsel R. R., Murray R. W., Rosenblum W. D., Khan S. A. Purification of pT181-encoded repC protein required for the initiation of plasmid replication. J Biol Chem. 1985 Jul 15;260(14):8571–8577. [PubMed] [Google Scholar]
  14. Koepsel R. R., Murray R. W., Rosenblum W. D., Khan S. A. The replication initiator protein of plasmid pT181 has sequence-specific endonuclease and topoisomerase-like activities. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6845–6849. doi: 10.1073/pnas.82.20.6845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Koths K., Dressler D. Analysis of the phiX DNA replication cycle by electron microscopy. Proc Natl Acad Sci U S A. 1978 Feb;75(2):605–609. doi: 10.1073/pnas.75.2.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Marians K. J., Soeller W., Zipursky S. L. Maximal limits of the Escherichia coli replication factor Y effector site sequences in pBR322 DNA. J Biol Chem. 1982 May 25;257(10):5656–5662. [PubMed] [Google Scholar]
  17. McMacken R., Ueda K., Kornberg A. Migration of Escherichia coli dnaB protein on the template DNA strand as a mechanism in initiating DNA replication. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4190–4194. doi: 10.1073/pnas.74.10.4190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nomura N., Low R. L., Ray D. S. Identification of ColE1 DNA sequences that direct single strand-to-double strand conversion by a phi X174 type mechanism. Proc Natl Acad Sci U S A. 1982 May;79(10):3153–3157. doi: 10.1073/pnas.79.10.3153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Primrose S. B., Ehrlich S. D. Isolation of plasmid deletion Mutants and study of their instability. Plasmid. 1981 Sep;6(2):193–201. doi: 10.1016/0147-619x(81)90066-4. [DOI] [PubMed] [Google Scholar]
  20. Schaller H. The intergenic region and the origins for filamentous phage DNA replication. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):401–408. doi: 10.1101/sqb.1979.043.01.046. [DOI] [PubMed] [Google Scholar]
  21. Scheer-Abramowitz J., Gryczan T. J., Dubnau D. Origin and mode of replication of plasmids pE194 and pUB110. Plasmid. 1981 Jul;6(1):67–77. doi: 10.1016/0147-619x(81)90054-8. [DOI] [PubMed] [Google Scholar]
  22. Shlomai J., Kornberg A. An Escherichia coli replication protein that recognizes a unique sequence within a hairpin region in phi X174 DNA. Proc Natl Acad Sci U S A. 1980 Feb;77(2):799–803. doi: 10.1073/pnas.77.2.799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Smits M. A., Schoenmakers J. G., Konings R. N. Expression of bacteriophage M13 DNA in vivo. Isolation, identification and characterization of phage-specific mRNA species. Eur J Biochem. 1980 Nov;112(2):309–321. doi: 10.1111/j.1432-1033.1980.tb07206.x. [DOI] [PubMed] [Google Scholar]
  24. Staudenbauer W. L., Scherzinger E., Lanka E. Replication of the colicin E1 plasmid in extracts of Escherichia coli: uncoupling of leading strand from lagging strand synthesis. Mol Gen Genet. 1979;177(1):113–120. doi: 10.1007/BF00267260. [DOI] [PubMed] [Google Scholar]
  25. Sutcliffe J. G. pBR322 restriction map derived from the DNA sequence: accurate DNA size markers up to 4361 nucleotide pairs long. Nucleic Acids Res. 1978 Aug;5(8):2721–2728. doi: 10.1093/nar/5.8.2721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Twigg A. J., Sherratt D. Trans-complementable copy-number mutants of plasmid ColE1. Nature. 1980 Jan 10;283(5743):216–218. doi: 10.1038/283216a0. [DOI] [PubMed] [Google Scholar]
  27. Van der Ende A., Teertstra R., Weisbeek P. J. n' Protein activator sites of plasmid pBR322 are not essential for its DNA replication. J Mol Biol. 1983 Jul 5;167(3):751–756. doi: 10.1016/s0022-2836(83)80108-9. [DOI] [PubMed] [Google Scholar]
  28. Veltkamp E., Stuitje A. R. Replication and structure of the bacteriocinogenic plasmids Clo DF13 and CoI E1. Plasmid. 1981 Jan;5(1):76–99. doi: 10.1016/0147-619x(81)90078-0. [DOI] [PubMed] [Google Scholar]
  29. Zipursky S. L., Marians K. J. Escherichia coli factor Y sites of plasmid pBR322 can function as origins of DNA replication. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6111–6115. doi: 10.1073/pnas.78.10.6111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zipursky S. L., Marians K. J. Identification of two Escherichia coli factor Y effector sites near the origins of replication of the plasmids (ColE1 and pBR322. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6521–6525. doi: 10.1073/pnas.77.11.6521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. van der Ende A., Teertstra R., van der Avoort H. G., Weisbeek P. J. Initiation signals for complementary strand DNA synthesis on single-stranded plasmid DNA. Nucleic Acids Res. 1983 Jul 25;11(14):4957–4975. doi: 10.1093/nar/11.14.4957. [DOI] [PMC free article] [PubMed] [Google Scholar]

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