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Expression of recA gene of Deinococcus radiodurans in Escherichia coli cells

  • Genetics of Microorganisms
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Abstract

Plasmids pKS5 and pKSrec30 carrying normal and mutant alleles of the Deinococcus recA gene controlled by the lactose promoter slightly increase radioresistance of Escherichia coli cells with mutations in genes recA and ssb. The RecA protein of D. radiodurans is expressed in E. coli cells, and its synthesis can be supplementary induced. The radioprotective effect of the xenologic protein does not exceed 1.5 fold and yields essentially to the contribution of plasmid pUC19-recA1.1 harboring the E. coli recA + gene in the recovery of resistance of the ΔrecA deletion mutant. These data suggest that the expression of D. radiodurans recA gene in E. coli cells does not complement mutations at gene recA in the chromosome possibly due to structural and functional peculiarities of the D. radiodurans RecA protein.

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References

  1. Moseley, B.E.B. and Copland, H.J.R., Isolation and Properties of a Recombination-Deficient Mutant of Micrococcus radiodurans, J. Bacteriol., 1975, vol. 121, no. 2, pp. 422–428.

    PubMed  CAS  Google Scholar 

  2. Gutman, P.D., Carroll, J.D., Masters, C.I., et al., Sequencing, Targeted Mutagenesis and Expression of a recA Gene Required for the Extreme Radioresistance of Deinococcus radiodurans, Gene, 1994, vol. 141, no. 1, pp. 31–37.

    Article  PubMed  CAS  Google Scholar 

  3. Verbenko, V.N., Kuznetsova, L.V., Krup’yan, E.P., and Suslov, A.V., Operator-Constitutive Mutation in recA Gene Enhances Radiation Resistance of Escherishia coli, Russ. J. Genet., 2009, vol. 45, no. 8, (in press).

  4. Makarova, K.S., Aravind, L., Wolf, Y.I., et al., Genome of the Extremely Radiation-Resistant Bacterium Deinococcus radiodurans Viewed from the Perspective of Comparative Genomics, Microbiol. Mol. Biol. Rev., 2001, vol. 65, no. 1, pp. 44–79.

    Article  PubMed  CAS  Google Scholar 

  5. White, O., Eisen, J.A., Heidelberg, J.F., et al., Genome Sequence of the Radioresistant Bacterium Deinococcus radiodurans, Science, 1999, vol. 286, no. 5444, pp. 1671–1577.

    Google Scholar 

  6. Narumi, I., Satoh, K., Kikuchi, M., et al., The LexA Protein from Deinococcus radiodurans Is not Involved in RecA Induction Following Gamma Irradiation, J. Bacteriol., 2001, vol. 183, no. 23, pp. 6951–6956.

    Article  PubMed  CAS  Google Scholar 

  7. Earl, A.M., Mohundro, M.M., Mian, I.S., et al., The IrrE Protein of Deinococcus radiodurans R1 Is a Novel Regulator of recA Expression, J. Bacteriol., 2002, vol. 184, no. 22, pp. 6216–6224.

    Article  PubMed  CAS  Google Scholar 

  8. Narumi, I., Satoh, K., Kikuchi, M., et al., Molecular Analysis of the Deinococcus radiodurans recA Locus and Identification of a Mutation Site in a DNA Repair-Deficient Mutant, rec30, Mutat. Res., 1999, vol. 435, no. 3, pp. 233–243.

    PubMed  CAS  Google Scholar 

  9. Alexseyev, A.A., Bakhlanova, I.V., Zaitsev, E.N., et al., 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 

  10. Sambrook, J., Fritsch, E.F., and Maniatis, T., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor: Cold Spring Harbor Lab., 1989.

    Google Scholar 

  11. Verbenko, V.N., Smol’nikova, A.V., and Kalinin, V.L., Effect of a Null Mutation in the priA Gene on the Radiation Resistance of Escherichia coli, Russ. J. Genet., 2000, vol. 36, no. 3, pp. 318–321.

    CAS  Google Scholar 

  12. Kim, J.-I., Sharma, A.K., Abbott, S.N., et al., RecA Protein from the Extremely Radioresistant Bacterium Deinococcus radiodurans: Expression, Purification, and Characterization, J. Bacteriol., 2002, vol. 184, no. 6, pp. 1649–1660.

    Article  PubMed  CAS  Google Scholar 

  13. Roca, A.I. and Cox, M.M., RecA Protein: Structure, Function, and Role in Recombinational DNA Repair, Prog. Nucleic Acid Res. Mol. Biol., 1997, vol. 56, pp. 129–223.

    Article  PubMed  CAS  Google Scholar 

  14. Ogawa, T., Wabiko, H., Tsurimoto, T., et al., Characteristics of Purified RecA Protein and the Regulation of Its Synthesis in vivo, Cold Spring Harb. Symp. Quant. Biol., 1979, vol. 43, pp. 909–915.

    PubMed  CAS  Google Scholar 

  15. Kowalczykowski, S.C., Clow, J., and Soman, R., et al., Effects of the Escherichia coli SSB Protein on the Binding of Escherichia coli RecA Protein to Single-Stranded DNA: Demonstration of Competitive Binding and the Lack of a Specific Protein-Protein Interaction, J. Mol. Biol., 1987, vol. 193, no. 1, pp. 81–95.

    Article  PubMed  CAS  Google Scholar 

  16. Kowalczykowski, S.C. and Krupp, R.A., Effects of Escherichia coli SSB Protein on the Single-Stranded DNA-Dependent ATPase Activity of Escherichia coli RecA Protein: Evidence That SSB Protein Facilitates the Binding of RecA Protein to Regions of Secondary Structure within Single-Stranded DNA, J. Mol. Biol., 1987, vol. 193, no. 1, pp. 97–113.

    Article  PubMed  CAS  Google Scholar 

  17. Lavery, P.E. and Kowalczykowski, S.C., A Postsynaptic Role for Single-Stranded DNA-Binding Protein in RecA Protein-Promoted DNA Strand Exchange, J. Biol. Chem., 1992, vol. 267, no. 13, pp. 9315–9320.

    PubMed  CAS  Google Scholar 

  18. Bernstein, D.A., Eggington, J.M., Killoran, M.P., et al., Crystal Structure of the Deinococcus radiodurans Single-Stranded DNA-Binding Protein Suggests a Mechanism for Coping with DNA Damage, Proc. Natl. Acad. Sci. USA, 2004, vol. 101, no. 23, pp. 8575–8580.

    Article  PubMed  CAS  Google Scholar 

  19. Egginton, J.M., Haruta, N., Wood, E.A., et al., The Single-Stranded DNA-Binding Protein of Deinococcus radiodurans, BMC Microbiol., 2004, vol. 4, no. 2.

  20. Kim, J.I. and Cox, M.M., The RecA Proteins of Deinococcus radiodurans and Escherichia coli Promote DNA Strand Exchange via Inverse Pathways, Proc. Natl. Acad. Sci. USA, 2002, vol. 99, no. 12, pp. 7917–7921.

    Article  PubMed  CAS  Google Scholar 

  21. Blattner, F.R., Plunkett, G., III, Bloch, C.A., et al., The Complete Genome Sequence of Escherichia coli K-12, Science, 1997, vol. 277, no. 5331, pp. 1453–1474.

    Article  PubMed  CAS  Google Scholar 

  22. Glassberg, J., Meyer, R.R., and Kornberg, A., Mutant Single-Strand Binding Protein of Escherichia coli: Genetic and Physiological Characterization, J. Bacteriol., 1979, vol. 140, no. 1, pp. 14–19.

    PubMed  CAS  Google Scholar 

  23. Nair, S. and Steyn, L.M., Cloning and Expression in Escherichia coli of a recA Homologue from Mycobacterium tuberculosis, J. Gen. Microbiol., 1991, vol. 137, no. 10, pp. 2409–2414.

    PubMed  CAS  Google Scholar 

  24. Kokjohn, T.A. and Miller, R.V., Characterization of the Pseudomonas aeruginosa recA Analog and Its Protein Product: rec-102 Is a Mutant Allele of the P. aeruginosa PAO recA Gene, J. Bacteriol., 1987, vol. 169, no. 4, pp. 1499–1508.

    PubMed  CAS  Google Scholar 

  25. Kuhl, S.A., Zimmer, J.A., and Rohatgi, P., Complementation of Bacteriophage Induction and Recombination Defects in Escherichia coli RecA − Mutants by Expression of the Cloned T4 Bacteriophage uvsX Gene, Curr. Microbiol., 2003, vol. 46, no. 2, pp. 88–93.

    Article  PubMed  CAS  Google Scholar 

  26. Cox, M.M., The Bacterial RecA Protein: Structure, Function, and Regulation, Topics in Current Genetics, New York: Springer, 2007, pp. 53–94.

    Google Scholar 

  27. Ishimori, K., Sommer, S., Bailone, A., et al., Characterization of a Mutant RecA Protein That Facilitates Homologous Genetic Recombination but not Recombinational DNA Repair: RecA423, J. Mol. Biol., 1996, vol. 264, no. 4, pp. 696–712.

    Article  PubMed  CAS  Google Scholar 

  28. Eggler, A.L., Lusetti, S.L., and Cox, M.M., The C-Terminus of the E. coli RecA Protein Modulates the DNA Binding Competition with SSB, J. Biol. Chem., 2003, vol. 278, no. 18, pp. 16389–16396.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. St. Petersburg Konstantinov Institute of Nuclear Physics, Russian Academy of Sciences, Gatchina, 188300, Russia

    V. N. Verbenko, L. V. Kuznetsova, E. P. Krupyan & V. I. Shalguev

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  1. V. N. Verbenko
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  2. L. V. Kuznetsova
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  3. E. P. Krupyan
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  4. V. I. Shalguev
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Correspondence to V. N. Verbenko.

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Original Russian Text © V.N. Verbenko, L.V. Kuznetsova, E.P. Krupyan, V.I. Shalguev, 2009, published in Genetika, 2009, Vol. 45, No. 10, pp. 1353–1360.

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Verbenko, V.N., Kuznetsova, L.V., Krupyan, E.P. et al. Expression of recA gene of Deinococcus radiodurans in Escherichia coli cells. Russ J Genet 45, 1192–1199 (2009). https://doi.org/10.1134/S1022795409100068

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  • DOI: https://doi.org/10.1134/S1022795409100068

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