1887

Abstract

Two-component regulatory systems have been widely implicated in bacterial virulence. To investigate the role of one such system in , a strain was constructed in which the system was deleted by homologous recombination. The mutant strain (Tame15) showed a growth defect after infection of macrophages and was attenuated in both immunodeficient and immunocompetent mice. Competitive hybridization of total RNA from the wild-type and mutant strains to a whole-genome microarray was used to identify changes in gene expression resulting from the deletion. One operon was highly up-regulated in the mutant, indicating that probably has a role as a repressor of this operon. Other genes which were up- or down-regulated were also identified. Many of the genes showing down-regulation are involved in normal growth of the bacterium, indicating that the mutant strain is subject to some type of growth slow-down or stress. Genes showing differential expression were further grouped according to their pattern of gene expression under other stress conditions. From this analysis 50 genes were identified which are the most likely to be controlled by RegX3. Most of these genes are of unknown function and no obvious motifs were found upstream of the genes identified. Thus, it has been demonstrated that the two-component system is involved in the virulence of and a number of genes controlled by this system have been identified.

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2003-06-01
2019-11-17
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References

  1. Banu, S., Honore, N., Saint-Joanis, B., Philpott, D., Prevost, M. C. & Cole, S. T. ( 2002; ). Are the PE-PGRS proteins of Mycobacterium tuberculosis variable surface antigens? Mol Microbiol 44, 9–19.[CrossRef]
    [Google Scholar]
  2. Barrett, J. F. & Hoch, J. A. ( 1998; ). Two-component signal transduction as a target for microbial anti-infective therapy. Antimicrob Agents Chemother 42, 1529–1536.
    [Google Scholar]
  3. Belisle, J. T. & Sonnenberg, M. G. ( 1998; ). Isolation of genomic DNA from mycobacteria. In Mycobacteria Protocols, pp. 31–44. Edited by T. Parish & N. G. Stoker. Totowa, NJ: Humana Press.
  4. Betts, J. C., Lukey, P. T., Robb, L. C., McAdam, R. A. & Duncan, K. ( 2002; ). Evaluation of a nutrient starvation model of Mycobacterium tuberculosis persistence by gene and protein expression profiling. Mol Microbiol 43, 717–731.[CrossRef]
    [Google Scholar]
  5. Cole, S. T., Brosch, R., Parkhill, J. & 39 other authors ( 1998; ). Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature 393, 537–544.[CrossRef]
    [Google Scholar]
  6. Dussurget, O., Timm, J., Gomez, M., Gold, B., Yu, S. W., Sabol, S. Z., Holmes, R. K., Jacobs, W. R. & Smith, I. ( 1999; ). Transcriptional control of the iron-responsive fxbA gene by the mycobacterial regulator IdeR. J Bacteriol 181, 3402–3408.
    [Google Scholar]
  7. Dziejman, M. & Mekalanos, J. J. ( 1995; ). Two-component signal transduction and its role in the expression of bacterial virulence factors. In Two-component Signal Transduction, pp. 305–317. Edited by J. A. Hoch & T. J. Silhavy. Washington, DC: American Society for Microbiology.
  8. Ewann, F., Jackson, M., Pethe, K., Cooper, A., Mielcarek, N., Ensergueix, D., Gicquel, B., Locht, C. & Supply, P. ( 2002; ). Transient requirement of the PrrA–PrrB two-component system for early intracellular multiplication of Mycobacterium tuberculosis. Infect Immun 70, 2256–2263.[CrossRef]
    [Google Scholar]
  9. Feng, Z. Y., Caceres, N. E., Sarath, G. & Barletta, R. G. ( 2002; ). Mycobacterium smegmatis l-alanine dehydrogenase (Ald) is required for proficient utilization of alanine as a sole nitrogen source and sustained anaerobic growth. J Bacteriol 184, 5001–5010.[CrossRef]
    [Google Scholar]
  10. Garcia Vescovi, E., Soncini, F. C. & Groisman, E. A. ( 1996; ). Mg2+ as an extracellular signal: environmental regulation of Salmonella virulence. Cell 84, 165–174.[CrossRef]
    [Google Scholar]
  11. Groisman, E. A. & Heffron, F. ( 1995; ). Regulation of Salmonella virulence by two-component regulatory systems. In Two-component Signal Transduction, pp. 319–332. Edited by J. A. Hoch & T. J. Silhavy. Washington, DC: American Society for Microbiology.
  12. Himpens, S., Locht, C. & Supply, P. ( 2000; ). Molecular characterization of the mycobacterial SenX3–RegX3 two-component system: evidence for autoregulation. Microbiology 146, 3091–3098.
    [Google Scholar]
  13. Hoch, J. A. ( 2000; ). Two-component and phosphorelay signal transduction. Curr Opin Microbiol 3, 165–170.[CrossRef]
    [Google Scholar]
  14. Lee, T. I., Rinaldi, N. J., Robert, F. & 18 other authors ( 2002; ). Transcriptional regulatory networks in Saccharomyces cerevisiae. Science 298, 799–804.[CrossRef]
    [Google Scholar]
  15. Lukey, P. T. ( 2001; ). Macrophage virulence assays. In Mycobacterium tuberculosis Protocols, pp. 271–280. Edited by T. Parish & N. G. Stoker. Totowa, NJ: Humana Press.
  16. Manganelli, R., Voskuil, M. I., Schoolnik, G. K. & Smith, I. ( 2001; ). The Mycobacterium tuberculosis ECF sigma factor sigma(E): role in global gene expression and survival in macrophages. Mol Microbiol 41, 423–437.[CrossRef]
    [Google Scholar]
  17. Manganelli, R., Voskuil, M. I., Schoolnik, G. K., Dubnau, E., Gomez, M. & Smith, I. ( 2002; ). Role of the extracytoplasmic-function sigma factor sigma(H) in Mycobacterium tuberculosis global gene expression. Mol Microbiol 45, 365–374.[CrossRef]
    [Google Scholar]
  18. Movahedzadeh, F., Gonzalez-Y-Merchand, J. A. & Cox, R. A. ( 2001; ). Transcription start site mapping. In Mycobacterium tuberculosis Protocols, pp. 105–124. Edited by T. Parish & N. G. Stoker. Totowa, NJ: Humana Press.
  19. Parish, T. & Stoker, N. G. ( 2000; ). Use of a flexible cassette method to generate a double unmarked Mycobacterium tuberculosis tlyA plcABC mutant by gene replacement. Microbiology 146, 1969–1975.
    [Google Scholar]
  20. Parish, T., Turner, J. & Stoker, N. G. ( 2001; ). amiA is a negative regulator of acetamidase expression in Mycobacterium smegmatis. BMC Microbiol 1, 19.[CrossRef]
    [Google Scholar]
  21. Parish, T., Smith, D. A., Kendall, S., Casali, N., Bancroft, G. J. & Stoker, N. G. ( 2003; ). Deletion of two-component regulatory systems increases virulence of Mycobacterium tuberculosis. Infect Immun 71, 1134–1140.[CrossRef]
    [Google Scholar]
  22. Perez, E., Samper, S., Bordas, Y., Guilhot, C., Gicquel, B. & Martin, C. ( 2001; ). An essential role for phoP in Mycobacterium tuberculosis virulence. Mol Microbiol 41, 179–187.[CrossRef]
    [Google Scholar]
  23. Ramakrishnan, L., Federspiel, N. A. & Falkow, S. ( 2000; ). Granuloma-specific expression of Mycobacterium virulence proteins from the glycine-rich PE-PGRS family. Science 288, 1436–1439.[CrossRef]
    [Google Scholar]
  24. Rodriguez, G. M., Voskuil, M. I., Gold, B., Schoolnik, G. K. & Smith, I. ( 2002; ). ideR, an essential gene in Mycobacterium tuberculosis: role of IdeR in iron-dependent gene expression, iron metabolism, and oxidative stress response. Infect Immun 70, 3371–3381.[CrossRef]
    [Google Scholar]
  25. Sherman, D. R., Voskuil, M., Schnappinger, D., Liao, R. L., Harrell, M. I. & Schoolnik, G. K. ( 2001; ). Regulation of the Mycobacterium tuberculosis hypoxic response gene encoding alpha-crystallin. Proc Natl Acad Sci U S A 98, 7534–7539.[CrossRef]
    [Google Scholar]
  26. Smith, D. A., Parish, T., Stoker, N. G. & Bancroft, G. J. ( 2001; ). Characterization of auxotrophic mutants of Mycobacterium tuberculosis and their potential as vaccine candidates. Infect Immun 69, 1142–1150.[CrossRef]
    [Google Scholar]
  27. Stewart, G. R., Wernisch, L., Stabler, R., Mangan, J. A., Hinds, J., Laing, K. G., Young, D. B. & Butcher, P. D. ( 2002; ). Dissection of the heat-shock response in Mycobacterium tuberculosis using mutants and microarrays. Microbiology 148, 3129–3138.
    [Google Scholar]
  28. Urao, T., Yamaguchi-Shinozaki, K. & Shinozaki, K. ( 2000; ). Two-component systems in plant signal transduction. Trends Plant Sci 5, 67–74.[CrossRef]
    [Google Scholar]
  29. Wayne, L. G. & Sohaskey, C. D. ( 2001; ). Nonreplicating persistence of Mycobacterium tuberculosis. Annu Rev Microbiol 55, 139–163.[CrossRef]
    [Google Scholar]
  30. Wernisch, L., Soneji, S., Wietzorrek, A., Parish, T., Hinds, J., Butcher, P. D. & Stoker, N. G. ( 2003; ). Analysis of whole-genome microarray replicates using mixed models. Bioinformatics 19, 53–61.[CrossRef]
    [Google Scholar]
  31. Wren, B. W., Colby, S. M., Cubberley, R. R. & Pallen, M. J. ( 1992; ). Degenerate PCR primers for the amplification of fragments from genes encoding response regulators from a range of pathogenic bacteria. FEMS Microbiol Lett 99, 287–292.[CrossRef]
    [Google Scholar]
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