Skip to main content
Log in

Operator-constitutive mutation in the recA gene enhances radiation resistance of Escherichia coli

  • Genetics of Microorganisms
  • Published:
Russian Journal of Genetics Aims and scope Submit manuscript

Abstract

Plasmid pUC19-recAoc carrying a mutant allele of the recA gene, which plays the key role in the control of the SOS repair system and homologous recombinational repair, causes a 1.5-fold increase in radiation resistance of Escherichia coli ΔrecA cells, as compared to the wild-type recA + cells. The protective effect of this plasmid is drastically reduced in mutant lexA3 recAΔ21 deficient in the LexA protein and in induction of the SOS regulon. Plasmid pUC19-recAoc effectively suppresses UV sensitivity of the ΔrecA mutant. Mutation recAo20 allows constitutive high-level synthesis of the RecA protein. This mutation impairs the SOS box in the operator site of the recA gene and enhances heterology of the dimer LexA binding site. These data confirm that high level of the RecA protein synthesis per se is not sufficient for the expression of γ-inducible functions and that the derepression of lexA-dependent genes, other than recA gene, is necessary for the complete induction of the SOS repair system.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Friedberg, E.C., Walker, G.C., and Siede, W., DNA Repair and Mutagenesis, Washington, DC: ASM Press, 1995.

    Google Scholar 

  2. Bresler, S.E., Verbenko, V.N., and Kalinin, V.L., Escherichia coli K-12 Mutants with Enhanced Resistance to Ionizing Radiation: II. Study of DNA Damage and Repair in Gamma-Irradiated Cells, Genetika (Moscow), 1982, vol. 18, no. 8, pp. 1245–1254.

    CAS  Google Scholar 

  3. Verbenko, V.N. and Kalinin, V.L., Growth of Radiation Resistance in Bacteriophages as a Result of Intensifying Expression of the Stress System in Host Cells, Russ. J. Genet., 1995, vol. 31, no. 12, pp. 1386–1392.

    CAS  Google Scholar 

  4. Pollard, E.C. and Randall, E.P., Studies on the Inducible Inhibitor of Radiation-Induced DNA Degradation of Escherichia coli, Radiat. Res., 1973, vol. 55, no. 2, pp. 265–279.

    Article  PubMed  CAS  Google Scholar 

  5. McGrew, D.A. and Knight, K.L., Molecular Design and Functional Organization of the RecA Protein, Crit. Rev. Biochem. Mol. Biol., 2003, vol. 38, pp. 385–432.

    Article  PubMed  CAS  Google Scholar 

  6. Karu, A.E. and Belk, E.D., Induction of E. coli recA Protein via recBC and Alternative Pathways: Quantitation by Enzyme-Linked Immunosorbent Assay (ELISA), Mol. Gen. Genet., 1982, vol. 185, no. 2, pp. 275–282.

    Article  PubMed  CAS  Google Scholar 

  7. Salles, B. and Paoletti, C., Control of UV Induction of RecA Protein, Proc. Natl. Acad. Sci. USA, 1983, vol. 80, no. 1, pp. 65–69.

    Article  PubMed  CAS  Google Scholar 

  8. Volkert, M.R., Margossian, L.J., and Clark, A.J., Evidence That rnmB Is the Operator of the Escherichia coli recA Gene, Proc. Natl. Acad. Sci. USA, 1981, vol. 78, no. 3, pp. 1786–1780.

    Article  PubMed  CAS  Google Scholar 

  9. Ginsburg, H., Edmiston, S.H., Harper, J., and Mount, D.W., Isolation and Characterization of an Operator-Constitutive Mutation in the recA Gene of E. coli K-12, Mol. Gen. Genet., 1982, vol. 187, no. 1, pp. 4–11.

    Article  PubMed  CAS  Google Scholar 

  10. Alexseyev, A.A., Bakhlanova, I.V., Zaitsev, E.N., and Lanzov, V.A., Genetic Characteristics of New recA Mutants of Escherichia coli K-12, J. Bacteriol., 1996, vol. 178, no. 7, pp. 2018–2024.

    PubMed  CAS  Google Scholar 

  11. Sambrook, J., Fritsch, E.F., and Maniatis, T., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Lab., 1989, 2nd ed.

    Google Scholar 

  12. Verbenko, V.N., Kuznetsova, L.V., and Kalinin, V.L., Mutant Alleles for Radioresistance from the Escherichia coli Strain Gamr444: Cloning and Preliminary Characteristics, Russ. J. Genet., 1994, vol. 30, no. 6, pp. 756–762.

    CAS  Google Scholar 

  13. Verbenko, V.N., Smol’nikova, A.V., and Kalinin, V.L., Effect of a Null Mutation in the priA Gene on Radioresistance of Escherichia coli, Russ. J. Genet., 2000, vol. 36, no. 3, pp. 245–248.

    CAS  Google Scholar 

  14. Laemmli, U.K. and Favre, M., Maturation of the Head of Bacteriophage T4: I. DNA Packaging Events, J. Mol. Biol., 1973, vol. 80, no. 4, pp. 575–599.

    Article  PubMed  CAS  Google Scholar 

  15. Walker, G.C., Mutagenesis and Inducible Response to Deoxyribonucleic Acid Damage in Escherichia coli, Microbiol. Rev., 1984, vol. 48, no. 1, pp. 60–93.

    PubMed  CAS  Google Scholar 

  16. Verbenko, V.N., Kuznetsova, L.V., Smol’nikova, A.V., and Kalinin, V.L., Effect of Insertional Mutation in the cspA Gene Encoding the Major Cold-Shock Protein on Radiation Resistance of Escherichia coli, Russ. J. Genet., 2003, vol. 39, no. 6, pp. 618–626.

    Article  CAS  Google Scholar 

  17. Bresler, S.E., Verbenko, V.N., and Kalinin, V.L., Escherichia coli K-12 Mutants with Enhanced Resistance to Ionizing Radiation, III. The Effect of rec and lexA Mutations on Radioresistance, Genetika (Moscow), 1984, vol. 20, no. 5, pp. 746–755.

    CAS  Google Scholar 

  18. Lewis, L.K., Harlow, G.R., Gregg-Jolly, L.A., et al., Identification of High Affinity Binding Sites for LexA Which Define New DNA Damage-Inducible Genes in Escherichia coli, J. Mol. Biol., 1994, vol. 241, no. 4, pp. 507–523.

    Article  PubMed  CAS  Google Scholar 

  19. Courcelle, J., Khodursky, A., Peter, B., et al., Comparative Gene Expression Profiles Following UV Exposure in Wild-Type and SOS-Deficient Escherichia coli, Genetics, 2001, vol. 158, no. 1, pp. 41–64.

    PubMed  CAS  Google Scholar 

  20. Picksley, S.M., Attfield, P.V., and Lloyd, R.G., Repair of DNA Double-Strand Breaks in Escherichia coli K12 Requires a Functional recN Product, Mol. Gen. Genet., 1984, vol. 195, nos. 1–2, pp. 267–274.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to V. N. Verbenko.

Additional information

Original Russian Text © V.N. Verbenko, L.V. Kuznetsova, E.P. Krupyan, A.V. Suslov, 2009, published in Genetika, 2009, Vol. 45, No. 8, pp. 1048–1054.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Verbenko, V.N., Kuznetsova, L.V., Krupyan, E.P. et al. Operator-constitutive mutation in the recA gene enhances radiation resistance of Escherichia coli . Russ J Genet 45, 917–923 (2009). https://doi.org/10.1134/S1022795409080043

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S1022795409080043

Keywords

Navigation