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Abstract

Nitrate reduction by is regulated by control of the transport of nitrate into the cell by NarK2. When oxygen was introduced into hypoxic cultures, nitrite production was quickly inhibited. The nitrate-reducing enzyme itself is relatively insensitive to oxygen, suggesting that the inhibition of nitrite production by oxygen was a result of interference with nitrate transport. This was not due to degradation of NarK2, as the inhibition was reversed by the removal of oxygen although chloramphenicol prevented new synthesis of NarK2. The oxidant potassium ferricyanide was added to anaerobic cultures to produce a positive redox potential in the absence of oxygen. Nitrite production decreased, signifying that oxidizing conditions, rather than oxygen itself, were responsible for the inhibition of nitrate transport. Nitric oxide added to cultures allowed NarK2 to be active even in the presence of oxygen. A similar result was obtained with hydroxylamine and ethanol, both of which interfere with oxygen utilization and the electron transport chain. It is proposed that NarK2 senses the redox state of the cell, possibly by monitoring the flow of electrons to cytochrome oxidase, and adjusts its activity so that nitrate is transported under reducing, but not under oxidizing, conditions.

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2005-11-01
2019-11-21
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References

  1. Adams, L. B., Dinauer, M. C., Morgenstern, D. E. & Krahenbuhl, J. L. ( 1997; ). Comparison of the roles of reactive oxygen and nitrogen intermediates in the host response to Mycobacterium tuberculosis using transgenic mice. Tuber Lung Dis 78, 237–246.[CrossRef]
    [Google Scholar]
  2. Alefounder, P. R., McCarthy, J. E. G. & Ferguson, S. J. ( 1981; ). The basis of the control of nitrate reduction by oxygen in Paracoccus denitrificans. FEMS Microbiol Lett 12, 321–326.[CrossRef]
    [Google Scholar]
  3. Alefounder, P. R., Greenfield, A. J., McCarthy, J. E. G. & Ferguson, S. J. ( 1983; ). Selection and organisation of denitrifying electron-transfer pathways in Paracoccus denitrificans. Biochim Biophys Acta 724, 20–39.[CrossRef]
    [Google Scholar]
  4. Boshoff, H. I. M. & Barry, C. E. I. ( 2005; ). Tuberculosis – metabolism and respiration in the absence of growth. Nature Rev 3, 70–80.
    [Google Scholar]
  5. Brunori, M., Giuffre, A., Forte, E., Mastronicola, D., Barone, M. C. & Sarti, P. ( 2004; ). Control of cytochrome c oxidase activity by nitric oxide. Biochim Biophys Acta 1655, 365–371.[CrossRef]
    [Google Scholar]
  6. Chan, J., Tanaka, K., Carroll, D., Flynn, J. & Bloom, B. R. ( 1995; ). Effects of nitric oxide synthase inhibitors on murine infection with Mycobacterium tuberculosis. Infect Immun 63, 736–740.
    [Google Scholar]
  7. Chan, E. D., Chan, J. & Schluger, N. W. ( 2001; ). What is the role of nitric oxide in murine and human host defense against tuberculosis? Am J Respir Cell Mol Biol 25, 606–612.[CrossRef]
    [Google Scholar]
  8. Clementi, E., Brown, G. C., Foxwell, N. & Moncada, S. ( 1999; ). On the mechanism by which vascular endothelial cells regulate their oxygen consumption. Proc Natl Acad Sci U S A 96, 1559–1562.[CrossRef]
    [Google Scholar]
  9. 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]
  10. Denis, K. S., Dias, F. M. & Rowe, J. J. ( 1990; ). Oxygen regulation of nitrate transport by diversion of electron flow in Escherichia coli. J Biol Chem 265, 18095–18097.
    [Google Scholar]
  11. Dolin, P. J., Raviglione, M. C. & Kochi, A. ( 1994; ). Global tuberculosis incidence and mortality during 1990–2000. Bull W H O 72, 213–220.
    [Google Scholar]
  12. Fritz, C., Maass, S., Kreft, A. & Bange, F.-C. ( 2002; ). Dependence of Mycobacterium bovis BCG on anaerobic nitrate reductase for persistence is tissue specific. Infect Immun 70, 286–291.[CrossRef]
    [Google Scholar]
  13. Goldstein, S., Russo, A. & Sammuni, A. ( 2004; ). Reactions of PTIO and carboxy-PTIO with NO, NO2, and O2. J Biol Chem 278, 50949–50955.
    [Google Scholar]
  14. Hernandez, D. & Rowe, J. J. ( 1988; ). Oxygen inhibition of nitrate uptake is a general regulatory mechanism in nitrate respiration. J Biol Chem 263, 7937–7939.
    [Google Scholar]
  15. John, P. ( 1977; ). Aerobic and anaerobic bacterial respiration monitored by electrodes. J Gen Microbiol 98, 231–238.[CrossRef]
    [Google Scholar]
  16. Kayser, E.-B., Hoppel, C. L., Morgan, P. G. & Sedensky, M. M. ( 2003; ). A mutation in mitochondrial complex I increases ethanol sensitivity in Caenorhabditis elegans. Alcohol Clin Exp Res 27, 584–592.[CrossRef]
    [Google Scholar]
  17. Kendall, S. L., Movahedzadeh, F., Rison, S. C. G., Wernisch, L., Parish, T., Duncan, K., Betts, J. C. & Stoker, N. G. ( 2004; ). The Mycobacterium tuberculosis dosRS two-component system is induced by multiple stresses. Tuberculosis 84, 247–255.[CrossRef]
    [Google Scholar]
  18. Kucera, I. & Skladal, P. ( 1990; ). Hydroxylamine as an inhibitor and terminal acceptor in the respiratory chain of the bacterium Paracoccus denitrificans. Gen Physiol Biophys 9, 501–518.
    [Google Scholar]
  19. Kucera, I., Karlovský, P. & Dadák, V. ( 1981; ). Control of nitrate respiration in Paracoccus denitrificans by oxygen. FEMS Microbiol Lett 12, 391–394.[CrossRef]
    [Google Scholar]
  20. Kucera, I., Kaplan, P. & Zeman, A. ( 1996; ). Oxygen increases the steady-state level of nitrate in denitrifying cells of Paracoccus denitrificans. FEMS Microbiol Lett 145, 163–166.[CrossRef]
    [Google Scholar]
  21. MacMicking, J. D., North, R. J., LaCourse, R., Mudgett, J. S., Shah, S. K. & Nathan, C. F. ( 1997; ). Identification of nitric oxide synthase as a protective locus against tuberculosis. Proc Natl Acad Sci U S A 94, 5243–5248.[CrossRef]
    [Google Scholar]
  22. Nathan, C. ( 2002; ). Inducible nitric oxide synthase in the tuberculous human lung. Am J Resp Critic Care Med 166, 130–131.[CrossRef]
    [Google Scholar]
  23. Noji, S. & Taniguchi, S. ( 1987; ). Molecular oxygen controls nitrate transport of Escherichia coli nitrate-respiring cells. J Biol Chem 262, 9441–9443.
    [Google Scholar]
  24. Ohno, H., Zhu, G., Mohan, V. P., Chu, D., Kohno, S., Jacobs, W. R. J. & Chan, J. ( 2003; ). The effects of reactive nitrogen intermediates on gene expression in Mycobacterium tuberculosis. Cell Microbiol 5, 637–648.[CrossRef]
    [Google Scholar]
  25. Sherman, D. R., Voskuil, M., Schnappinger, D., Liao, R., Harrell, M. I. & Schoolnik, G. K. ( 2001; ). Regulation of the Mycobacterium tuberculosis hypoxic response gene encoding α-crystallin. Proc Natl Acad Sci U S A 98, 7534–7539.[CrossRef]
    [Google Scholar]
  26. Shi, L., Sohaskey, C. D., Kana, B. D., Dawes, S., North, R. J., Mizrahi, V. & Gennaro, M. L. ( 2005; ). Changes in energy metabolism in Mycobacterium tuberculosis as revealed by bacterial transcriptional profiling during mouse lung infection and under in vitro conditions affecting aerobic respiration. Proc Natl Acad Sci U S A. In press.
    [Google Scholar]
  27. Sohaskey, C. D. & Wayne, L. G. ( 2003; ). Role of narK2X and narGHJI in hypoxic upregulation of nitrate reduction by Mycobacterium tuberculosis. J Bacteriol 185, 7247–7256.[CrossRef]
    [Google Scholar]
  28. Unden, G., Trageser, M. & Duchene, A. ( 1990; ). Effect of positive redox potentials (>+400 mV) on the expression of anaerobic respiratory enzymes in Escherichia coli. Mol Microbiol 4, 315–319.[CrossRef]
    [Google Scholar]
  29. Voskuil, M. I., Schnappinger, D., Harrell, M. I., Visconti, K. C., Dolganov, G. M., Sherman, D. R. & Schoolnik, G. K. ( 2002; ). Inhibition of respiration by nitric oxide induces a Mycobacterium tuberculosis persistence program. J Exp Med 198, 705–713.
    [Google Scholar]
  30. Wade, M. M. & Zhang, Y. ( 2004; ). Anaerobic incubation conditions enhance pyrazinamide activity against Mycobacterium tuberculosis. J Med Microbiol 53, 769–773.[CrossRef]
    [Google Scholar]
  31. Wayne, L. G. & Doubek, J. R. ( 1965; ). Classification and identification of Mycobacteria. II Tests employing nitrate and nitrite as substrate. Am Rev Resp Dis 91, 738–745.
    [Google Scholar]
  32. Wayne, L. G. & Hayes, L. G. ( 1996; ). An in vitro model for sequential study of shiftdown of Mycobacterium tuberculosis through two stages of nonreplicating persistence. Infect Immun 64, 2062–2069.
    [Google Scholar]
  33. Wayne, L. G. & Hayes, L. G. ( 1999; ). Nitrate reduction as a marker for hypoxic shiftdown of Mycobacterium tuberculosis. Tuber Lung Dis 79, 127–132.
    [Google Scholar]
  34. Wayne, L. G. & Sohaskey, C. D. ( 2001; ). Nonreplicating persistence of Mycobacterium tuberculosis. Annu Rev Microbiol 55, 139–163.[CrossRef]
    [Google Scholar]
  35. Weber, I., Fritz, C., Ruttkowski, S., Kreft, A. & Bange, F.-C. ( 2000; ). Anaerobic nitrate reductase (narGHJI) activity of Mycobacterium bovis BCG in vitro and its contribution to virulence in immunodeficient mice. Mol Microbiol 35, 1017–1025.[CrossRef]
    [Google Scholar]
  36. Zhang, Y., Wade, M. M., Scorpio, A., Zhang, H. & Sun, Z. ( 2003; ). Mode of action of pyrazinamide: disruption of Mycobacterium tuberculosis membrane transport and energetics by pyrazinoic acid. J Antimicrob Chemother 52, 790–795.[CrossRef]
    [Google Scholar]
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