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. 1991 Oct 11;19(19):5379–5383. doi: 10.1093/nar/19.19.5379

Probing the activation of the replicative origin of broad host-range plasmid R1162 with Tus, the E.coli anti-helicase protein.

H S Zhou 1, C Byrd 1, R J Meyer 1
PMCID: PMC328902  PMID: 1923822

Abstract

The E.coli Tus protein is an anti-helicase involved in the termination of chromosome replication. The binding site for this protein, ter, was cloned into derivatives of the broad host-range plasmid R1162. The ter site caused the orientation-specific termination of plasmid replication fork movement in cell extracts containing Tus. Plasmids were constructed so that two sites for initiation of R1162 replication flanked the iteron-containing domain of the origin. In these plasmids, the site next to the AT-rich region within the iteron-containing domain was more active. In addition, when ter was placed between the more active site and the iterons, initiation of replication from this site was specifically inhibited. The data support a model for entry of the essential, plasmid-encoded helicase at one side of the direct repeats, and for its movement primarily in one direction away from these repeats to activate the initiation sites for DNA replication.

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Selected References

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

  1. Bramhill D., Kornberg A. A model for initiation at origins of DNA replication. Cell. 1988 Sep 23;54(7):915–918. doi: 10.1016/0092-8674(88)90102-x. [DOI] [PubMed] [Google Scholar]
  2. Hershfield V., Boyer H. W., Yanofsky C., Lovett M. A., Helinski D. R. Plasmid ColEl as a molecular vehicle for cloning and amplification of DNA. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3455–3459. doi: 10.1073/pnas.71.9.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Hidaka M., Akiyama M., Horiuchi T. A consensus sequence of three DNA replication terminus sites on the E. coli chromosome is highly homologous to the terR sites of the R6K plasmid. Cell. 1988 Nov 4;55(3):467–475. doi: 10.1016/0092-8674(88)90033-5. [DOI] [PubMed] [Google Scholar]
  4. Hill T. M., Henson J. M., Kuempel P. L. The terminus region of the Escherichia coli chromosome contains two separate loci that exhibit polar inhibition of replication. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1754–1758. doi: 10.1073/pnas.84.7.1754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hill T. M., Marians K. J. Escherichia coli Tus protein acts to arrest the progression of DNA replication forks in vitro. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2481–2485. doi: 10.1073/pnas.87.7.2481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hill T. M., Pelletier A. J., Tecklenburg M. L., Kuempel P. L. Identification of the DNA sequence from the E. coli terminus region that halts replication forks. Cell. 1988 Nov 4;55(3):459–466. doi: 10.1016/0092-8674(88)90032-3. [DOI] [PubMed] [Google Scholar]
  7. Honda Y., Sakai H., Komano T., Bagdasarian M. RepB' is required in trans for the two single-strand DNA initiation signals in oriV of plasmid RSF1010. Gene. 1989 Aug 1;80(1):155–159. doi: 10.1016/0378-1119(89)90261-8. [DOI] [PubMed] [Google Scholar]
  8. Horiuchi T., Hidaka M. Core sequence of two separable terminus sites of the R6K plasmid that exhibit polar inhibition of replication is a 20 bp inverted repeat. Cell. 1988 Aug 12;54(4):515–523. doi: 10.1016/0092-8674(88)90073-6. [DOI] [PubMed] [Google Scholar]
  9. Khatri G. S., MacAllister T., Sista P. R., Bastia D. The replication terminator protein of E. coli is a DNA sequence-specific contra-helicase. Cell. 1989 Nov 17;59(4):667–674. doi: 10.1016/0092-8674(89)90012-3. [DOI] [PubMed] [Google Scholar]
  10. Lee E. H., Kornberg A., Hidaka M., Kobayashi T., Horiuchi T. Escherichia coli replication termination protein impedes the action of helicases. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9104–9108. doi: 10.1073/pnas.86.23.9104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lin L. S., Meyer R. J. DNA synthesis is initiated at two positions within the origin of replication of plasmid R1162. Nucleic Acids Res. 1987 Oct 26;15(20):8319–8331. doi: 10.1093/nar/15.20.8319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. MacAllister T., Khatri G. S., Bastia D. Sequence-specific and polarized replication termination in vitro: complementation of extracts of tus- Escherichia coli by purified Ter protein and analysis of termination intermediates. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2828–2832. doi: 10.1073/pnas.87.7.2828. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Meyer R., Hinds M., Brasch M. Properties of R1162, a broad-host-range, high-copy-number plasmid. J Bacteriol. 1982 May;150(2):552–562. doi: 10.1128/jb.150.2.552-562.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  15. Pelletier A. J., Hill T. M., Kuempel P. L. Location of sites that inhibit progression of replication forks in the terminus region of Escherichia coli. J Bacteriol. 1988 Sep;170(9):4293–4298. doi: 10.1128/jb.170.9.4293-4298.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Scherzinger E., Bagdasarian M. M., Scholz P., Lurz R., Rückert B., Bagdasarian M. Replication of the broad host range plasmid RSF1010: requirement for three plasmid-encoded proteins. Proc Natl Acad Sci U S A. 1984 Feb;81(3):654–658. doi: 10.1073/pnas.81.3.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Scherzinger E., Haring V., Lurz R., Otto S. Plasmid RSF1010 DNA replication in vitro promoted by purified RSF1010 RepA, RepB and RepC proteins. Nucleic Acids Res. 1991 Mar 25;19(6):1203–1211. doi: 10.1093/nar/19.6.1203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sista P. R., Mukherjee S., Patel P., Khatri G. S., Bastia D. A host-encoded DNA-binding protein promotes termination of plasmid replication at a sequence-specific replication terminus. Proc Natl Acad Sci U S A. 1989 May;86(9):3026–3030. doi: 10.1073/pnas.86.9.3026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Stougaard P., Molin S. Vertical dye-buoyant density gradients for rapid analysis and preparation of plasmid DNA. Anal Biochem. 1981 Nov 15;118(1):191–193. doi: 10.1016/0003-2697(81)90177-9. [DOI] [PubMed] [Google Scholar]
  20. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  21. Zhou H. S., Meyer R. J. Deletion of sites for initiation of DNA synthesis in the origin of broad host-range plasmid R1162. J Mol Biol. 1990 Aug 5;214(3):685–697. doi: 10.1016/0022-2836(90)90286-u. [DOI] [PubMed] [Google Scholar]
  22. de Massy B., Béjar S., Louarn J., Louarn J. M., Bouché J. P. Inhibition of replication forks exiting the terminus region of the Escherichia coli chromosome occurs at two loci separated by 5 min. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1759–1763. doi: 10.1073/pnas.84.7.1759. [DOI] [PMC free article] [PubMed] [Google Scholar]

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