Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1983 Dec;80(23):7132–7136. doi: 10.1073/pnas.80.23.7132

Identification of a primosome assembly site in the region of the ori 2 replication origin of the Escherichia coli mini-F plasmid.

R Imber, R L Low, D S Ray
PMCID: PMC390007  PMID: 6316346

Abstract

A primosome assembly site for F plasmid DNA replication has been identified. This site, which we term rriA (F), is localized to one strand of a 385-base-pair Sau3A restriction fragment very close to ori 2 and within the 2.25-kilobase DNA sequence required for replication and incompatibility of the entire F plasmid. rriA (F) was isolated by cloning into the deletion phage vector M13 delta Elac. This phage forms very faint plaques due to a deletion of the M13 complementary strand origin but forms large wild-type plaques when DNA single-strand initiation determinants are inserted. The single-stranded viral DNA of the Sau3A F-M13 delta Elac recombinant provides an effector site of dATP hydrolysis by the primosomal protein n'. It also provides an assembly site for the Escherichia coli primosome protein complex that directs the in vitro conversion of the single-stranded DNA to a double-stranded form by the same mechanism as that used by phi X174. Homologies of the nucleotide sequence between this F DNA sequence and the previously identified primosome assembly sites in phi X174 phage DNA and in ColE1 plasmid DNA (rriA and rriB) have been found. The sequences 5' G-T-G-A-G-C-G 3' and 5' G-N-G-G-A-A-G-C 3' or variations of these sequences occur from two to five times within each assembly locus. In addition, two distinct 15-base-pair sequences in rriA (F) are perfectly homologous to corresponding sequences in rriA (ColE1).

Full text

PDF

Images in this article

7134Image
on p.7134

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. Bouché J. P., Zechel K., Kornberg A. 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. [PubMed] [Google Scholar]
  4. Geider K., Kornberg A. Conversion of the M13 viral single strand to the double-stranded replicative forms by purified proteins. J Biol Chem. 1974 Jul 10;249(13):3999–4005. [PubMed] [Google Scholar]
  5. Hines J. C., Ray D. S. Construction and characterization of new coliphage M13 cloning vectors. Gene. 1980 Nov;11(3-4):207–218. doi: 10.1016/0378-1119(80)90061-x. [DOI] [PubMed] [Google Scholar]
  6. Hirota Y., Ryter A., Jacob F. Thermosensitive mutants of E. coli affected in the processes of DNA synthesis and cellular division. Cold Spring Harb Symp Quant Biol. 1968;33:677–693. doi: 10.1101/sqb.1968.033.01.077. [DOI] [PubMed] [Google Scholar]
  7. Kahn M. L., Figurski D., Ito L., Helinski D. R. 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. doi: 10.1101/sqb.1979.043.01.015. [DOI] [PubMed] [Google Scholar]
  8. Kim M. H., Hines J. C., Ray D. S. Viable deletions of the M13 complementary strand origin. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6784–6788. doi: 10.1073/pnas.78.11.6784. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lane H. E. Replication and incompatibility of F and plasmids in the IncFI Group. Plasmid. 1981 Jan;5(1):100–126. doi: 10.1016/0147-619x(81)90079-2. [DOI] [PubMed] [Google Scholar]
  10. Low R. L., Arai K., Kornberg A. Conservation of the primosome in successive stages of phi X174 DNA replication. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1436–1440. doi: 10.1073/pnas.78.3.1436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. McDonell M. W., Simon M. N., Studier F. W. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol. 1977 Feb 15;110(1):119–146. doi: 10.1016/s0022-2836(77)80102-2. [DOI] [PubMed] [Google Scholar]
  13. Murotsu T., Matsubara K., Sugisaki H., Takanami M. Nine unique repeating sequences in a region essential for replication and incompatibility of the mini-F plasmid. Gene. 1981 Nov;15(2-3):257–271. doi: 10.1016/0378-1119(81)90135-9. [DOI] [PubMed] [Google Scholar]
  14. Nishimura Y., Caro L., Berg C. M., Hirota Y. Chromosome replication in Escherichia coli. IV. Control of chromosome replication and cell division by an integrated episome. J Mol Biol. 1971 Feb 14;55(3):441–456. doi: 10.1016/0022-2836(71)90328-7. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Nomura N., Low R. L., Ray D. S. Selective cloning of Co1E1 DNA initiation sequences using the cloning vector M13 delta E101. Gene. 1982 Jun;18(3):239–246. doi: 10.1016/0378-1119(82)90161-5. [DOI] [PubMed] [Google Scholar]
  17. Nomura N., Ray D. S. Expression of a DNA strand initiation sequence of ColE1 plasmid in a single-stranded DNA phage. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6566–6570. doi: 10.1073/pnas.77.11.6566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ray D. S., Hines J. C., Kim M. H., Imber R., Nomura N. M13 vectors for selective cloning of sequences specifying initiation of DNA synthesis on single-stranded templates. Gene. 1982 Jun;18(3):231–238. doi: 10.1016/0378-1119(82)90160-3. [DOI] [PubMed] [Google Scholar]
  19. Schekman R., Weiner A., Kornberg A. Multienzyme systems of DNA replication. Science. 1974 Dec 13;186(4168):987–993. doi: 10.1126/science.186.4168.987. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Watson L. A., Phua S. H., Bergquist P. L., Lane H. E. An Mr 29000 protein is essential for mini-F maintenance in E. coli. Gene. 1982 Sep;19(2):173–178. doi: 10.1016/0378-1119(82)90003-8. [DOI] [PubMed] [Google Scholar]
  22. 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]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES