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Russian Journal of Genetics

, Volume 43, Issue 1, pp 35–40 | Cite as

Initiator protein DnaA of Escherichia coli is a negative replication regulator of linear phage-plasmid N15

  • A. V. Mardanov
  • N. V. Ravin
Genetics of Microorganisms
  • 40 Downloads

Abstract

Temperate bacteriophage N15 in the lysogenic state is incapable of integrating in the chromosome of Escherichia coli and represents a linear plasmid with covalently closed ends. The phage repA gene, the product of which possesses activities of primase and helicase, ensures replication of N15 DNA. The ori site of initiation of N15 replication contains binding sites for RepA and a potential site of binding the bacterial initiator protein DnaA. It was shown in our work that replication of miniplasmids based on N15 replicon as well as replication of N15 DNA during lytic growth do not depend on DnaA. Moreover, introducing mutations into the potential DnaA binding site increases the copy number of circular and linear miniplasmids that contain repA gene. These data suggest that DnaA is a negative rather than positive regulator of phage N15 replication. This is assumed to be caused by properties of interaction between RepA and DnaA during initiation of N15 replication or by transcriptional silencing of repA gene due to the binding of DnaA to the ori site located within the repA coding sequence.

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References

  1. 1.
    Watson, J., Origin of Concatemeric T7 DNA, Nature New Biol., 1972, vol. 239, pp. 197–201.PubMedCrossRefGoogle Scholar
  2. 2.
    Ravin, N.V., Mechanisms of Replication and Telomere Resolution of the Linear Plasmid Prophage N15, FEMS Microbiol. Letts., 2003, vol. 221, pp. 1–6.CrossRefGoogle Scholar
  3. 3.
    Svarchevskii, A.N. and Rybchin, V.N., Physical Mapping of N15 Plasmid DNA, Mol. Genet. Mikrobiol. Virusol., 1984, vol. 10, pp. 16–22.Google Scholar
  4. 4.
    Ravin, N.V., Kuprianov, V.V., Gilcrease, E.B., and Casjens, S.R., Bidirectional Replication from an Internal Ori Site of the Linear N15 Plasmid Prophage, Nucleic Acids Res., 2003, vol. 31, pp. 6552–6560.PubMedCrossRefGoogle Scholar
  5. 5.
    Lobocka, M.B., Svarchevsky, A.N., Rybchin, V.N., and Yarmolinsky, M.B., Characterization of the Primary Immunity Region of the Escherichia coli Linear Plasmid Prophage N15, J. Bacteriol., 1996, vol. 178, pp. 2902–2910.PubMedGoogle Scholar
  6. 6.
    Mardanov, A.V. and Ravin, N.V., Functional Characterization of the repA Replication Gene of Linear Plasmid Prophage N15, Res. Microbiol, 2006, vol. 157, pp. 176–183.PubMedCrossRefGoogle Scholar
  7. 7.
    Svarchevskii, A.N. and Rybchin, V.N., Characteristic of Bacteriophage N15 Plasmid Features, Mol. Genet. Mikrobiol. Virusol., 1984, vol. 10, pp. 34–39.Google Scholar
  8. 8.
    Tilly, K., Independence of Bacteriophage N15 Lytic and Linear Plasmid Replication from the Heat Shock Proteins DnaJ, DnaK, and GrpE, J. Bacteriol., 1991, vol. 173, pp. 6639–6642.PubMedGoogle Scholar
  9. 9.
    Messer, W., The Bacterial Replication Initiator DnaA. DnaA and OriC, the Bacterial Mode to Initiate DNA Replication, FEMS Microbiol. Rev., 2002, vol. 26, pp. 355–374.PubMedGoogle Scholar
  10. 10.
    Hansen, E.B. and Yarmolinsky, M.B., Host Participation in Plasmid Maintenance: Dependence Upon dnaA of Replications Derived from P1 and F, Proc. Natl. Acad. Sci. USA, 1986, vol. 83, pp. 4423–4427.PubMedCrossRefGoogle Scholar
  11. 11.
    Schaper, S. and Messer, W., Interaction of the Initiator Protein DnaA of Escherichia coli with Its DNA Target, J. Biol. Chem., 1995, vol. 270, pp. 17 622–17 626.Google Scholar
  12. 12.
    Sasaki, I. and Bertani, G., Growth Abnormalities in Hfr Derivatives of Escherichia coli Strain C, J. Gen. Microbiol., 1965, vol. 40, pp. 365–376.PubMedGoogle Scholar
  13. 13.
    Grant, S.G., Jessee, J., Bloom, F.R., and Hanahan, D., Differential Plasmid Rescue from Transgenic Mouse DNAs Into Escherichia coli Methylation-Restriction Mutants, Proc. Natl. Acad. Sci. USA, 1990, vol. 87, pp. 4645–4649.PubMedCrossRefGoogle Scholar
  14. 14.
    Tocchetti, A., Galimberti, G., Deho, G., and Ghisotti, D., Characterization of the oril and oriII Origins of Replication of Phage-Plasmid P4, J. Virol., 1998, vol. 73, pp. 7308–7316.Google Scholar
  15. 15.
    Filip, C.C., Allen, J.S., Gustafson, R.A., et al., Bacterial Cell Division Regulation: Characterization of the dnaH Locus of Escherichia coli, J. Bacteriol., 1974, vol. 119, pp. 443–449.PubMedGoogle Scholar
  16. 16.
    Ravin, N.V., Rech, J., and Lane, D., Mapping of Functional Domains in F Plasmid Partition Proteins Reveals a Bipartite SopB-Recognition Domain in SopA, J. Mol. Biol., 2003, vol. 329, pp. 875–889.PubMedCrossRefGoogle Scholar
  17. 17.
    Ravin, N.V., Svarchevsky, A.N., and Deho, G., The Antiimunity System of Phage-Plasmid N15: Identification of the Antirepressor Gene and Its Control by a Small Processed RNA, Mol. Microbiol., 1999, vol. 34, pp. 980–994.PubMedCrossRefGoogle Scholar
  18. 18.
    Messer, W. and Weigel, C., DnaA Initiator Also a Transcription Factor, Mol. Microbiol., 1997, vol. 24, pp. 1–6.PubMedCrossRefGoogle Scholar
  19. 19.
    Mukhopadhyay, G., Carr, K.M., Kaguni, J.M., and Chattoraj, D.K., OPen-Complex Formation by the Host Initiator, DnaA, at the Origin of P1 Plasmid Replication, EMBO J., 1993, vol. 12, pp. 4547–4544.PubMedGoogle Scholar
  20. 20.
    Bernander, R., Dasgupta, S., and Nordstrom, K., The Escherichia coli Cell Cycle and the Plasmid R1 Replication Cycle in the Absence of the DnaA Protein, Cell, 1991, vol. 64, pp. 1145–1153.PubMedCrossRefGoogle Scholar
  21. 21.
    Masai, H. and Arai, K.I., RepA and DnaA Proteins Are Required for Initiation of R1 Plasmid Replication in Vitro and Interact with the oriR Sequence, Proc. Natl. Acad. Sci. USA, 1987, vol. 84, pp. 4781–4785.PubMedCrossRefGoogle Scholar
  22. 22.
    Ziegelin, G., Tegtmeyer, N., Lurz, R., et al., The repA Gene of the Linear Yersinia enterocolytica Prophage PY54 Functions as a Circular Minimal Replicon in Escherichia coli, J. Bacteriol., 2005, vol. 187, pp. 3445–3454.PubMedCrossRefGoogle Scholar
  23. 23.
    Ortega-Jimenez, S., Giraldo-Suarez, R., Fernandez-Tresguerres, M.E., et al., DnaA Dependent Replication of Plasmid R1 Occurs in the Presence of Point Mutations that Disrupt the DnaA Box of OriR, Nucleic Acids Res., 1992, vol. 20, pp. 2547–2551.PubMedCrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Inc. 2007

Authors and Affiliations

  • A. V. Mardanov
    • 1
  • N. V. Ravin
    • 1
  1. 1.Center of BioengineeringRussian Academy of SciencesMoscowRussia

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