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Persistence of Mycobacterium tuberculosis in macrophages and mice requires the glyoxylate shunt enzyme isocitrate lyase

Nature volume 406, pages 735–738 (2000)Cite this article

Abstract

Mycobacterium tuberculosis claims more human lives each year than any other bacterial pathogen. Infection is maintained in spite of acquired immunity and resists eradication by antimicrobials1,2. Despite an urgent need for new therapies targeting persistent bacteria, our knowledge of bacterial metabolism throughout the course of infection remains rudimentary. Here we report that persistence of M. tuberculosis in mice is facilitated by isocitrate lyase (ICL), an enzyme essential for the metabolism of fatty acids3,4. Disruption of the icl gene attenuated bacterial persistence and virulence in immune-competent mice without affecting bacterial growth during the acute phase of infection. A link between the requirement for ICL and the immune status of the host was established by the restored virulence of Δicl bacteria in interferon-γ knockout mice. This link was apparent at the level of the infected macrophage: Activation of infected macrophages increased expression of ICL, and the Δicl mutant was markedly attenuated for survival in activated but not resting macrophages. These data suggest that the metabolism of M. tuberculosis in vivo is profoundly influenced by the host response to infection, an observation with important implications for the treatment of chronic tuberculosis.

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Figure 1: Absence of ICL does not affect growth or stationary phase survival in vitro.
Figure 2: ICL is required for persistence and virulence of M. tuberculosis in immune-competent mice.
Figure 3: The level of ICL–GFP is higher in bacteria within activated macrophages.
Figure 4: The requirement for ICL is determined by the immune status of the host cell.
Figure 5: The Δicl mutant is lethal to immune-deficient mice.

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References

  1. McKinney, J. D., Jacobs, W. R. & Bloom, B. R. in Emerging Infections (eds Krause, R., Gallin, J. I. & Fauci, A. S.) 51–146 (Academic, New York, 1998).

    Book  Google Scholar 

  2. Parrish, N. M., Dick, J. D. & Bishai, W. R. Mechanisms of latency in Mycobacterium tuberculosis . Trends Microbiol. 6, 107– 112 (1998).

    Article  CAS  Google Scholar 

  3. Clark, D. P. & Cronan, J. E. Jr in Escherichia coli and Salmonella: Cellular and Molecular Biology (ed. Neidhardt, F. C.) 343–357 (ASM Press, Washington DC, 1996).

    Google Scholar 

  4. Cronan, J. E. Jr. & LaPorte, D. in Escherichia coli and Salmonella: Cellular and Molecular Biology (ed. Neidhardt, F. C.) 206–216 (ASM Press, Washington DC, 1996).

    Google Scholar 

  5. Segal, W. in The Mycobacteria: A Sourcebook (eds Kubica, G. P. & Wayne, L. G.) 547–573 (Dekker, New York, 1984 ).

    Google Scholar 

  6. Wheeler, P. R. & Ratledge, C. in Tuberculosis: Pathogenesis, Protection, and Control (ed. Bloom, B. R.) 353– 385 (ASM Press, Washington DC, 1994).

    Google Scholar 

  7. Höner zu Bentrup, K., Miczak, A., Swenson, D. L. & Russell, D. G. Characterization of activity and expression of isocitrate lyase in Mycobacterium avium and Mycobacterium tuberculosis. J. Bacteriol. 181, 7161–7167 ( 1999).

    Article  Google Scholar 

  8. Sturgill-Koszycki, S., Haddix, P. L. & Russell, D. G. The interaction between Mycobacterium and the macrophage analyzed by two-dimensional polyacrylamide gel electrophoresis. Electrophoresis 18, 2558– 2565 (1997).

    Article  CAS  Google Scholar 

  9. Graham, J. E. & Clark-Curtiss, J. E. Identification of Mycobacterium tuberculosis RNAs synthesized in response to phagocytosis by human macrophages by selective capture of transcribed sequences (SCOTS). Proc. Natl Acad. Sci. USA 96, 11554–11559 (1999).

    Article  ADS  CAS  Google Scholar 

  10. Farewell, A., Diez, A. A., DiRusso, C. C. & Nystrom, T. Role of the Escherichia coli FadR regulator in stasis survival and growth phase-dependent expression of the uspA, fad, and fab genes. J. Bacteriol. 178, 6443–6450 (1996).

    Article  CAS  Google Scholar 

  11. Spector, M. P. et al. The medium-/long-chain fatty acyl-CoA dehydrogenase (fadF) gene of Salmonella typhimurium is a phase 1 starvation-stress response (SSR) locus. Microbiol. 145, 15– 31 (1999).

    Article  CAS  Google Scholar 

  12. Suryanarayana Murthy, P., Sirsi, M. & Ramakrishnan, T. Effect of age on the enzymes of tricarboxylic acid and related cycles in Mycobacterium tuberculosis H37Rv. Amer. Rev. Resp. Dis. 108, 689–690 (1973).

    Google Scholar 

  13. Wayne, L. G. & Lin, K. Y. Glyoxylate metabolism and adaptation of Mycobacterium tuberculosis to survival under anaerobic conditions. Infect. Immunol. 37, 1042– 1049 (1982).

    Article  CAS  Google Scholar 

  14. Rees, R. J. W. & Hart, P. D. Analysis of the host-parasite equilibrium in chronic murine tuberculosis by total and viable bacillary counts. Br. J. Exp. Pathol. 42, 83–88 (1961).

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Wallace, J. G. The heat resistance of tubercle bacilli in the lungs of infected mice. Am. Rev. Respir. Dis. 83, 866–871 (1961).

    CAS  PubMed  Google Scholar 

  16. Hart, P. D. & Young, M. R. Interference with normal phagosome–lysosome fusion in macrophages, using ingested yeast cells and suramin. Nature 256, 47–49 ( 1975).

    Article  ADS  CAS  Google Scholar 

  17. Sturgill-Koszycki, S. et al. Lack of acidification in Mycobacterium phagosomes produced by exclusion of the vesicular proton-ATPase. Science 263, 678–681 (1994).

    Article  ADS  CAS  Google Scholar 

  18. Sturgill-Koszycki, S., Schaible, U. E. & Russell, D. G. Mycobacterium-containing phagosomes are accessible to early endosomes and reflect a transitional state in normal phagosome biogenesis. EMBO J. 15, 6960–6968 (1996).

    Article  CAS  Google Scholar 

  19. Clemens, D. L. & Horwitz, M. A. The Mycobacterium tuberculosis phagosome interacts with early endosomes and is accessible to exogenously administered transferrin. J. Exp. Med. 184, 1349–1355 (1996).

    Article  CAS  Google Scholar 

  20. James, P. E., Grinberg, O. Y., Michaels, G. & Swartz, H. M. Intraphagosomal oxygen in stimulated macrophages. J. Cell. Physiol. 163, 241–247 ( 1995).

    Article  CAS  Google Scholar 

  21. Schaible, U. E., Sturgill-Koszycki, S., Schlesinger, P. H. & Russell, D. G. Cytokine activation leads to acidification and increases maturation of Mycobacterium avium-containing phagosomes in murine macrophages. J. Immunol. 160, 1290–1296 (1998).

    CAS  PubMed  Google Scholar 

  22. Via, L. E. et al. Effects of cytokines on mycobacterial phagosome maturation. J. Cell Sci. 111, 897– 905 (1998).

    Article  CAS  Google Scholar 

  23. Pelicic, V. et al. Efficient allelic exchange and transposon mutagenesis in Mycobacterium tuberculosis. Proc Natl Acad Sci USA 94, 10955–10960 (1997).

    Article  ADS  CAS  Google Scholar 

  24. Mitchison, D. A. Treatment of tuberculosis. The Mitchell lecture 1979. J. R. Coll. Physicians Lond. 14, 91–99 (1980).

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Girling, D. J. in The Biology of the Mycobacteria, Volume 3: Clinical Aspects of Mycobacterial Disease (eds Ratledge, C., Stanford, J. & Grange, J. M.) 285 –323 (Academic, London, 1989).

    Google Scholar 

  26. Snapper, S. B., Melton, R. E., Mustafa, S., Kieser, T. & Jacobs, W. R. Jr Isolation and characterization of efficient plasmid transformation mutants of Mycobacterium smegmatis . Mol. Microbiol. 4, 1911– 1919 (1990).

    Article  CAS  Google Scholar 

  27. Stover, C. K. et al. New use of BCG for recombinant vaccines. Nature 351, 456–460 ( 1991).

    Article  ADS  CAS  Google Scholar 

  28. Pavelka, M. S. & Jacobs, W. R. Comparison of the construction of unmarked deletion mutants in Mycobacterium smegmatis , Mycobacterium bovis bacillus Calmette-Guerin, and Mycobacterium tuberculosis H37Rv by allellic exchange. J. Bacteriol. 181, 4780–4789 (1999).

    Article  CAS  Google Scholar 

  29. Cormack, B. P., Valdivia, R. H. & Falkow, S. FACS-optimized mutants of the green fluorescent protein (GFP). Gene 173, 33–38 (1996).

    Article  CAS  Google Scholar 

  30. Russo-Marie, F., Roederer, M., Sager, B., Herzenberg, L. A. & Kaiser, D. Beta-galactosidase activity in single differentiating bacterial cells. Proc Natl Acad Sci USA 90, 8194–8198 (1993).

    Article  ADS  CAS  Google Scholar 

  31. Sharma, V. et al. The structure of isocitrate lyase from Mycobacterium tuberculosis : a lynchpin to persistence of infection. Nature Struct. Biol. (in the press).

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Acknowledgements

J.D.M. and colleagues generated the data shown in Figs 1, 2, and 5. K.H.z.B. and colleagues generated the data shown in Figs 3 and 4. We thank B. R. Bloom for helpful discussions. J.D.M. was supported by a fellowship from the Helen Hay Whitney Foundation. J.D.M., E.J.M., W.T.C., K.H.z.B., A.M., D.S. and D.G.R. were supported by a grant from Glaxo Wellcome. All authors were supported by funds from the US Public Health Service.

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Author notes

  1. John D. McKinney & Wai-Tsing Chan

    Present address: The Rockefeller University, 1230 York Avenue, New York, New York, 10021, USA

  2. Kerstin Höner zu Bentrup, Dana Swenson & David G. Russell

    Present address: Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York, 14853, USA

  3. Andras Miczak

    Present address: Department of Microbiology, Albert Szent-Gyorgi Medical University, Szeged, POB 8-6701, Hungary

Authors and Affiliations

  1. Howard Hughes Medical Institute, Albert Einstein College of Medicine, Bronx, 10461, New York , USA

    John D. McKinney, Bing Chen & William R. Jacobs Jr

  2. Department of Molecular Microbiology Washington University School of Medicine, 660 S. Euclid Avenue , St. Louis, 63110, Missouri, USA

    Kerstin Höner zu Bentrup, Andras Miczak, Dana Swenson & David G. Russell

  3. The Rockefeller University, 1230 York Avenue, New York, 10021 , New York, USA

    Ernesto J. Muñoz-Elías & Wai-Tsing Chan

  4. Department of Biochemistry and Biophysics Texas A&M University, College Station, 77843, Texas, USA

    John D. McKinney, Kerstin Höner zu Bentrup, Andras Miczak, Dana Swenson, James C. Sacchettini & David G. Russell

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McKinney, J., zu Bentrup, K., Muñoz-Elías, E. et al. Persistence of Mycobacterium tuberculosis in macrophages and mice requires the glyoxylate shunt enzyme isocitrate lyase. Nature 406, 735–738 (2000). https://doi.org/10.1038/35021074

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