1887

Abstract

The complex includes , which causes tuberculosis in most mammals, including humans. In previous work, it was shown that ATCC 35721 has a mutation in its principal sigma factor gene, , causing a single amino acid change affecting binding of SigA with the accessory transcription factor WhiB3. ATCC 35721 is avirulent when inoculated subcutaneously into guinea pigs but can be restored to virulence by integration of wild-type to produce WAg320. Subsequently, it was surprising to discover that WAg320 was not virulent when inoculated intratracheally into the Australian brushtail possum (), a marsupial that is normally very susceptible to infection with . In this study, an complementation approach was used with ATCC 35721 to produce WAg322, which was virulent in possums, and to identify the virulence-restoring gene, . There are two point deletions in the gene of ATCC 35721 causing frameshift inactivation, one of which is also in the of BCG. Knockout of from ATCC 35723, a virulent strain of , produced WAg758, which was avirulent in both guinea pigs and possums, confirming that is a virulence gene. The effect on virulence of mode of infection versus animal species susceptibility was investigated by inoculating all the above strains by aerosol into guinea pigs and mice and comparing these to the earlier results. Characterization of PhoT indicated that it plays a role in phosphate uptake at low phosphate concentrations. At least , this role requires the presence of a wild-type gene and appears separate from the ability of to restore virulence to ATCC 35721. This study shows the advantages of using different animal models as tools for the molecular biological investigation of tuberculosis virulence.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.26469-0
2003-11-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/149/11/mic1493203.html?itemId=/content/journal/micro/10.1099/mic.0.26469-0&mimeType=html&fmt=ahah

References

  1. Aldwell F. E., Tucker I. G., de Lisle G. W., Buddle B. M. 2003; Oral delivery of Mycobacterium bovis BCG in a lipid formulation induces resistance to pulmonary tuberculosis in mice. Infect Immun 71:101–108
    [Google Scholar]
  2. Balasubramanian V., Wiegeshaus E. H., Smith D. W. 1992; Growth characteristics of recent sputum isolates of Mycobacterium tuberculosis in guinea pigs infected by the respiratory route. Infect Immun 60:4762–4767
    [Google Scholar]
  3. Banerjee S. K., Bhatt K., Misra P., Chakraborti P. K. 2000; Involvement of a natural transport system in the process of efflux-mediated drug resistance in Mycobacterium smegmatis. Mol Gen Genet 262:949–956
    [Google Scholar]
  4. Barry C. E. 2001; Interpreting cell wall ‘virulence factors' of Mycobacterium tuberculosis. Trends Microbiol 9:237–241
    [Google Scholar]
  5. Bhatt K., Banerjee S. K., Chakraborti P. K. 2000; Evidence that phosphate specific transporter is amplified in a fluoroquinolone resistant Mycobacterium smegmatis. Eur J Biochem 267:4028–4032
    [Google Scholar]
  6. Braibant M., Gilot P., Content J. 2000; The ATP binding cassette (ABC) transport systems of Mycobacterium tuberculosis. FEMS Microbiol Rev 24:449–467
    [Google Scholar]
  7. Cole S. T. 2002; Comparative and functional genomics of the Mycobacterium tuberculosis complex. Microbiology 148:2919–2928
    [Google Scholar]
  8. Cole S. T., Brosch R., Parkhill J. 38 other authors 1998; Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature 393:537–544
    [Google Scholar]
  9. Collins D. M., Kawakami R. P., de Lisle G. W., Pascopella L., Bloom B. R., Jacobs W. R. 1995; Mutation of the principal sigma factor causes loss of virulence in a strain of the Mycobacterium tuberculosis complex. Proc Natl Acad Sci U S A 92:8036–8040
    [Google Scholar]
  10. Collins D. M., Wilson T., Campbell S., Buddle B. M., Wards B. J., Hotter G., de Lisle G. W. 2002; Production of avirulent mutants of Mycobacterium bovis with vaccine properties by the use of illegitimate recombination and screening of stationary phase cultures. Microbiology 148:3019–3027
    [Google Scholar]
  11. Dannenberg A. M. Jr, Collins F. M. 2001; Progressive pulmonary tuberculosis is not due to increasing numbers of viable bacilli in rabbits, mice and guinea pigs, but is due to a continuous host response to mycobacterial products. Tuberculosis 81:229–242
    [Google Scholar]
  12. Darzins E. 1958 The Bacteriology of Tuberculosis Minneapolis, MN: University of Minnesota Press;
  13. de Lisle G. W., Mackintosh C. G., Bengis R. G. 2001; Mycobacterium bovis in free-living and captive wildlife, including farmed deer. Rev Sci Tech 20:86–111
    [Google Scholar]
  14. de Mendonca-Lima L., Bordat Y., Pivert E., Recchi C., Neyrolles O., Maitournam A., Gicquel B., Reyrat J. M. 2003; The allele encoding the mycobacterial Erp protein affects lung disease in mice. Cell Microbiol 5:65–73
    [Google Scholar]
  15. Fleischmann R. D., Alland D., Eisen J. A. 23 other authors 2002; Whole-genome comparison of Mycobacterium tuberculosis clinical and laboratory strains. J Bacteriol 184:5479–5490
    [Google Scholar]
  16. Garnier T., Eiglmeier K., Camus J-C. 19 other authors 2003; The complete genome sequence of Mycobacterium bovis. Proc Natl Acad Sci U S A 100:7877–7882
    [Google Scholar]
  17. Glickman M. S., Jacobs W. R. Jr 2001; Microbial pathogenesis of Mycobacterium tuberculosis: dawn of a discipline. Cell 104:477–485
    [Google Scholar]
  18. Higgins C. F. 2001; ABC transporters: physiology, structure and mechanism – an overview. Res Microbiol 152:205–210
    [Google Scholar]
  19. Jacobs W. R., Barrett J. F., Clark-Curtiss J. E., Curtiss R. III 1986; In vivo repackaging of recombinant cosmid molecules for analyses of Salmonella typhimurium, Streptococcus mutans, and mycobacterial genomic libraries. Infect Immun 52:101–109
    [Google Scholar]
  20. Kaushal D., Schroede B. G., Tyagi S. 8 other authors 2002; Reduced immunopathology and mortality despite tissue persistence in a Mycobacterium tuberculosis mutant lacking alternative sigma factor, SigH. Proc Natl Acad Sci U S A 99:8330–8335
    [Google Scholar]
  21. Kent P. T., Kubica G. P. 1985 Public Health Mycobacteriology, a Guide for the Level III Laboratory Atlanta, GA: US Department of Health and Human Service, Centres for Disease Control;
  22. Lewis K. N., Liao R., Guinn K. M., Hickey M. J., Smith S., Behr M. A., Sherman D. R. 2003; Deletion of RD1 from Mycobacterium tuberculosis mimics bacille Calmette-Guerin attenuation. J Infect Dis 187:117–123
    [Google Scholar]
  23. Linton K. J., Higgins C. F. 1998; The Escherichia coli ATP-binding cassette (ABC) proteins. Mol Microbiol 28:5–13
    [Google Scholar]
  24. McMurray D. N., Carlomagno M. A., Mintzer C. L., Tetzlaff C. L. 1985; Mycobacterium bovis BCG vaccine fails to protect protein-deficient guinea pigs against respiratory challenge with virulent Mycobacterium tuberculosis. Infect Immun 50:555–559
    [Google Scholar]
  25. North R. J., Ryan L., LaCource R., Mogues T., Goodrich M. E. 1999; Growth rate of mycobacteria in mice as an unreliable indication of mycobacterial virulence. Infect Immun 67:5483–5485
    [Google Scholar]
  26. Pascopella L., Collins F. M., Martin J. M., Lee M. H., Hatfull G. F., Stover C. K., Bloom B. R., Jacobs W. R. Jr 1994; Use of in vivo complementation in Mycobacterium tuberculosis to identify a genomic fragment associated with virulence. Infect Immun 62:1313–1319
    [Google Scholar]
  27. Pfeffer A., Buddle B. M., Aldwell F. E. 1994; Tuberculosis in the brushtail possum ( Trichosurus vulpecula) after intratracheal inoculation with a low dose of Mycobacterium bovis. J Comp Pathol 111:353–363
    [Google Scholar]
  28. Rao P. S. S., Lim T. M., Leung K. Y. 2003; Functional genomics approach to the identification of virulence genes involved in Edwardsiella tarda pathogenesis. Infect Immun 71:1343–1351
    [Google Scholar]
  29. Sarin J., Aggarwal S., Chaba R., Varshney G. C., Chakraborti P. K. 2001; B-subunit of phosphate-specific transporter from Mycobacterium tuberculosis is a thermostable ATPase. J Biol Chem 276:44590–44597
    [Google Scholar]
  30. Skinner M. A., Keen D. L., Parlane N. A., Yates G. F., Buddle B. M. 2002; Increased protection against bovine tuberculosis in the brushtail possum ( Trichosurus vulpecula) when BCG is administered with killed Mycobacterium vaccae. Tuberculosis 82:15–22
    [Google Scholar]
  31. Steyn A. J. C., Collins D. M., Hondalus M. K., Jacobs W. R., Kawakami R. P., Bloom B. R. 2002; Mycobacterium tuberculosis WhiB3 interacts with RpoV to affect host survival but is dispensable for in vivo growth. Proc Natl Acad Sci U S A 99:3147–3152
    [Google Scholar]
  32. van Soolingen D., Hermans P. W. M., de Haas P. E. W., Soll D. R., van Embden J. D. A. 1991; Occurrence and stability of insertion sequences in Mycobacterium tuberculosis complex strains: evaluation of an insertion-sequence dependent DNA polymorphism as a tool in the epidemiology of tuberculosis. J Clin Microbiol 29:2578–2586
    [Google Scholar]
  33. van Veen H. W. 1997; Phosphate transport in prokaryotes: molecules, mediators and mechanisms. Antonie van Leeuwenhoek 72:299–315
    [Google Scholar]
  34. von Kruger W. M., Humphreys S., Ketley J. M. 1999; A role for the PhoBR regulatory system homologue in the Vibrio cholerae phosphate-limitation response and intestinal colonization. Microbiology 145:2463–2475
    [Google Scholar]
  35. Wards B. J., Collins D. M. 1996; Electroporation at elevated temperatures substantially improves transformation efficiency of slow-growing mycobacteria. FEMS Microbiol Lett 145:101–105
    [Google Scholar]
  36. Wards B. J., de Lisle G. W., Collins D. M. 2000; An esat6 knockout mutant of Mycobacterium bovis produced by homologous recombination will contribute to the development of a live tuberculosis vaccine. Tubercle Lung Dis 80:185–189
    [Google Scholar]
  37. Wassenaar T. M., Gaastra W. 2001; Bacterial virulence: can we draw the line?. FEMS Microbiol Lett 201:1–7
    [Google Scholar]
  38. Wilson T. M., de Lisle G. W., Collins D. M. 1995; Effect of inhA and katG on isoniazid resistance and virulence of Mycobacterium bovis. Mol Microbiol 15:1009–1015
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.26469-0
Loading
/content/journal/micro/10.1099/mic.0.26469-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error