Slow induction of RecA by DNA damage in Free

Abstract

In mycobacteria, as in most bacterial species, the expression of RecA is induced by DNA damage. However, the authors show here that the kinetics of induction in and in are quite different: whilst maximum expression in occurred 3–6 h after addition of a DNA-damaging agent, incubation for 18–36 h was required to reach peak levels in . This is despite the fact that the promoter can be activated more rapidly when transferred to . In addition, it is demonstrated that in both species the DNA is sufficiently damaged to give maximum induction within the first hour of incubation with mitomycin C. The difference in the induction kinetics of between the two species was mirrored by a difference in the levels of DNA-binding-competent LexA following DNA damage. A decrease in the ability of LexA to bind to the SOS box was readily detected by 2 h in , whilst a decrease was not apparent until 18–24 h in and then only a very small decrease was observed.

Keyword(s): LexA , mycobacteria and SOS induction
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-147-12-3271
2001-12-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/147/12/1473271a.html?itemId=/content/journal/micro/10.1099/00221287-147-12-3271&mimeType=html&fmt=ahah

References

  1. Bertrand-Burggraf, E., Hurstel, S., Daune, M. & Schnarr, M. (1987). Promoter properties and negative regulation of the uvrA gene by the LexA repressor and its amino-terminal DNA binding domain. J Mol Biol 193, 293-302.[CrossRef] [Google Scholar]
  2. Brent, R. & Ptashne, M. (1981). Mechanism of action of the lexA gene product. Proc Natl Acad Sci USA 78, 4204-4208.[CrossRef] [Google Scholar]
  3. Casadaban, M. J., Martinez-Arias, A., Shapira, S. K. & Chou, J. (1983). Beta-galactosidase gene fusions for analyzing gene expression in Escherichia coli and yeast. Methods Enzymol 100, 293-308. [Google Scholar]
  4. Cheo, D. L., Bayles, K. W. & Yasbin, R. E. (1991). Cloning and characterization of DNA damage-inducible promoter regions from Bacillus subtilis. J Bacteriol 173, 1696-1703. [Google Scholar]
  5. Cheo, D. L., Bayles, K. W. & Yasbin, R. E. (1993). Elucidation of regulatory elements that control damage induction and competence induction of the Bacillus subtilis SOS system. J Bacteriol 175, 5907-5915. [Google Scholar]
  6. David, H. L. (1973). Response of mycobacteria to ultraviolet light radiation. Am Rev Respir Dis 108, 1175-1185. [Google Scholar]
  7. Davis, E. O., Jenner, P. J., Brooks, P. C., Colston, M. J. & Sedgwick, S. G. (1992). Protein splicing in the maturation of M. tuberculosis RecA protein: a mechanism for tolerating a novel class of intervening sequence. Cell 71, 201-210.[CrossRef] [Google Scholar]
  8. Demczuk, S., Harbers, M. & Vennstrom, B. (1993). Identification and analysis of all components of a gel retardation assay by combination with immunoblotting. Proc Natl Acad Sci USA 90, 2574-2578.[CrossRef] [Google Scholar]
  9. Durbach, S. I., Andersen, S. J. & Mizrahi, V. (1997). SOS induction in mycobacteria: analysis of the DNA-binding activity of a LexA-like repressor and its role in DNA damage induction of the recA gene from Mycobacterium smegmatis. Mol Microbiol 26, 643-653.[CrossRef] [Google Scholar]
  10. Fernandez de Henestrosa, A. R., Ogi, T., Aoyagi, S., Chafin, D., Hayes, J. J., Ohmori, H. & Woodgate, R. (2000). Identification of additional genes belonging to the LexA regulon in Escherichia coli. Mol Microbiol 35, 1560-1572. [Google Scholar]
  11. Friedberg, E., Walker, G. & Siede, W. (1995).DNA Repair and Mutagenesis. Washington, DC: American Society for Microbiology.
  12. Gassel, M. & Alonso, J. C. (1989). Expression of the recE gene during induction of the SOS response in Bacillus subtilis recombination-deficient strains. Mol Microbiol 3, 1269-1276.[CrossRef] [Google Scholar]
  13. Haijema, B. J., van Sinderen, D., Winterling, K., Kooistra, J., Venema, G. & Hamoen, L. W. (1996). Regulated expression of the dinR and recA genes during competence development and SOS induction in Bacillus subtilis. Mol Microbiol 22, 75-85.[CrossRef] [Google Scholar]
  14. Hiriyanna, K. T. & Ramakrishnan, T. (1986). Deoxyribonucleic acid replication time in Mycobacterium tuberculosis H37Rv. Arch Microbiol 144, 105-109.[CrossRef] [Google Scholar]
  15. Jacobs, W. R.Jr, Kalpana, G. V., Cirillo, J. D., Pascopella, L., Snapper, S. B., Udani, R. A., Jones, W., Barletta, R. G. & Bloom, B. R. (1991). Genetic systems for mycobacteria. Methods Enzymol 204, 537-555. [Google Scholar]
  16. Little, J. W. (1991). Mechanism of specific LexA cleavage: autodigestion and the role of RecA coprotease. Biochimie 73, 411-421.[CrossRef] [Google Scholar]
  17. Little, J. W. & Mount, D. W. (1982). The SOS regulatory system of Escherichia coli. Cell 29, 11-22.[CrossRef] [Google Scholar]
  18. Little, J. W., Mount, D. W. & Yanisch-Perron, C. R. (1981). Purified lexA protein is a repressor of the recA and lexA genes. Proc Natl Acad Sci USA 78, 4199-4203.[CrossRef] [Google Scholar]
  19. Lovett, C. M.Jr, Love, P. E., Yasbin, R. E. & Roberts, J. W. (1988). SOS-like induction in Bacillus subtilis: induction of the RecA protein analog and a damage-inducible operon by DNA damage in Rec+ and DNA repair-deficient strains. J Bacteriol 170, 1467-1474. [Google Scholar]
  20. Lovett, C. M.Jr, Cho, K. C. & O’Gara, T. M. (1993). Purification of an SOS repressor from Bacillus subtilis. J Bacteriol 175, 6842-6849. [Google Scholar]
  21. Miller, J. (1972).Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  22. Miller, M. C., Resnick, J. B., Smith, B. T. & Lovett, C. M.Jr (1996). The Bacillus subtilis dinR gene codes for the analogue of Escherichia coli LexA. Purification and characterization of the DinR protein. J Biol Chem 271, 33502-33508.[CrossRef] [Google Scholar]
  23. Movahedzadeh, F., Colston, M. J. & Davis, E. O. (1997a). Characterization of Mycobacterium tuberculosis LexA: recognition of a Cheo (Bacillus-type SOS) box. Microbiology 143, 929-936.[CrossRef] [Google Scholar]
  24. Movahedzadeh, F., Colston, M. J. & Davis, E. O. (1997b). Determination of DNA sequences required for regulated Mycobacterium tuberculosis RecA expression in response to DNA-damaging agents suggests that two modes of regulation exist. J Bacteriol 179, 3509-3518. [Google Scholar]
  25. Papavinasasundaram, K. G., Movahedzadeh, F., Keer, J. T., Stoker, N. G., Colston, M. J. & Davis, E. O. (1997). Mycobacterial recA is cotranscribed with a potential regulatory gene called recX. Mol Microbiol 24, 141-153.[CrossRef] [Google Scholar]
  26. Papavinasasundaram, K. G., Colston, M. J. & Davis, E. O. (1998). Construction and complementation of a recA deletion mutant of Mycobacterium smegmatis reveals that the intein in Mycobacterium tuberculosis recA does not affect RecA function. Mol Microbiol 30, 525-534.[CrossRef] [Google Scholar]
  27. Patel, B. K., Banerjee, D. K. & Butcher, P. D. (1991). Characterization of the heat shock response in Mycobacterium bovis BCG. J Bacteriol 173, 7982-7987. [Google Scholar]
  28. Raymond-Denise, A. & Guillen, N. (1992). Expression of the Bacillus subtilis dinR and recA genes after DNA damage and during competence. J Bacteriol 174, 3171-3176. [Google Scholar]
  29. Sambrook, J., Fritsch, E. & Maniatis, T. (1989).Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  30. Sassanfar, M. & Roberts, J. W. (1990). Nature of the SOS-inducing signal in Escherichia coli. The involvement of DNA replication. J Mol Biol 212, 79-96.[CrossRef] [Google Scholar]
  31. Snapper, S. B., Melton, R. E., Mustafa, S., Kieser, T. & Jacobs, W. R.Jr (1990). Isolation and characterization of efficient plasmid transformation mutants of Mycobacterium smegmatis. Mol Microbiol 4, 1911-1919.[CrossRef] [Google Scholar]
  32. Stover, C. K., de la Cruz, V. F., Fuerst, T. R. & 11 other authors (1991). New use of BCG for recombinant vaccines. Nature 351, 456–460.[CrossRef] [Google Scholar]
  33. Walker, G. C. (1984). Mutagenesis and inducible responses to deoxyribonucleic acid damage in Escherichia coli. Microbiol Rev 48, 60-93. [Google Scholar]
  34. Winterling, K. W., Levine, A. S., Yasbin, R. E. & Woodgate, R. (1997). Characterization of DinR, the Bacillus subtilis SOS repressor. J Bacteriol 179, 1698-1703. [Google Scholar]
  35. Winterling, K. W., Chafin, D., Hayes, J. J., Sun, J., Levine, A. S., Yasbin, R. E. & Woodgate, R. (1998). The Bacillus subtilis DinR binding site: redefinition of the consensus sequence. J Bacteriol 180, 2201-2211. [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-147-12-3271
Loading
/content/journal/micro/10.1099/00221287-147-12-3271
Loading

Data & Media loading...

Most cited Most Cited RSS feed