1887

Abstract

The mycobacterial cell wall consists of a core composed of peptidoglycan linked to the heteropolysaccharide arabinogalactan, which in turn is attached to mycolic acids. A variety of free lipids complements the mycolyl residues, whereas phosphatidylinositol mannosides (PIMs), lipoarabinomannan and proteins are interspersed in this framework. As a consequence, the cell envelope is extremely rich in lipids and early work has shown that the lipid content may vary with environmental conditions. To extend these studies, the influence of growth temperature on cell envelope components in , a temperature-resistant mycobacterial species, was investigated. Mycolic acid synthesis was reduced at 55 °C compared to 37 °C and the production of fatty acids, presumably precursors of mycolic acids, was increased. Since fatty acids are elongated by the type II fatty acid synthase complex and consequently by a mycobacterial β-ketoacyl acyl carrier protein synthase (KasA), leading to mycolic acids, the expression level of KasA was analysed by Western blotting. KasA expression was significantly decreased at 55 °C over 37 °C. Important changes in the mycolic acid composition were observed and characterized by reduced levels of cyclopropanation and the concomitant accumulation of the -olefin derivatives. In addition, striking differences involved in complex lipid composition, including acylated trehaloses and trehalose dimycolate (TDM) were also observed. At 55 °C, produced less TDM than at 37 °C, which could be explained by the down-regulation of antigen 85 (Ag85) expression as shown by Western blotting. The Ag85 complex represents a family of proteins known to catalyse the transfer of mycolates to trehalose, thereby generating TDM. Furthermore, at 55 °C the level of phosphatidyl-inositol hexamannoside (PIM) synthesis, but not that of other PIM species, was dramatically reduced. This observation could be correlated to a decrease of mannosyltransferase activity associated with membranes prepared from cells grown at 55 °C as compared to 37 °C. Altogether, this study suggests that mycobacteria are capable of inducing important cell-wall changes in response to temperature variations, which may represent a strategy developed by the bacteria to adapt to environmental changes.

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2002-10-01
2024-04-19
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References

  1. Apostolou I, Takahama Y, Belmant C. 9 other authors 1999; Murine natural killer T (NKT) cells contribute to the granulomatous reaction caused by mycobacterial cell walls. Proc Natl Acad Sci USA 96:5141–5146
    [Google Scholar]
  2. Baba T, Kaneda K, Kusunose E, Kusunose M., Yano I. 1989; Thermally adaptive changes of mycolic acids in Mycobacterium smegmatis . J Biochem 106:81–86
    [Google Scholar]
  3. Beatty W. L, Rhoades E. R, Ullrich H. J, Chatterjee D, Heuser J. E., Russell D. G. 2000; Trafficking and release of mycobacterial lipids from infected macrophages. Traffic 1:235–247
    [Google Scholar]
  4. Beatty W. L, Rhoades E. R, Hsu D. K, Liu F. T., Russell D. G. 2002; Association of a macrophage galactoside-binding protein with Mycobacterium -containing phagosomes. Cell Microbiol 4:167–176
    [Google Scholar]
  5. Bekierkunst A. 1968; Acute granulomatous response produced in mice by trehalose-6,6′-dimycolate. J Bacteriol 96:958–961
    [Google Scholar]
  6. Belisle J. T, Vissa V. D, Sievert T, Takayama K, Brennan P. J., Besra G. S. 1997; Role of the major antigen of Mycobacterium tuberculosis in cell wall biogenesis. Science 276:1420–1422
    [Google Scholar]
  7. Besra G. S. 1998; Preparation of cell-wall fractions from mycobacteria. Methods Mol Biol 101:91–107
    [Google Scholar]
  8. Besra G. S, Morehouse C. B, Rittner C. M, Waechter C. J., Brennan P. J. 1997; Biosynthesis of mycobacterial lipoarabinomannan. J Biol Chem 272:18460–18466
    [Google Scholar]
  9. Bloch H. 1950; Studies on the virulence of tubercle bacilli. Isolation and biological properties of a constituent of virulent organisms. J Exp Med 91:197–218
    [Google Scholar]
  10. Brennan P. J., Nikaido H. 1995; The envelope of mycobacteria. Annu Rev Biochem 64:29–63
    [Google Scholar]
  11. Camacho L. R, Constant P, Raynaud C, Lanéelle M. A, Triccas J. A, Gicquel B, Daffé M., Guilhot C. 2001; Analysis of the phthiocerol dimycocerosate locus of Mycobacterium tuberculosis . J Biol Chem 276:19845–19854
    [Google Scholar]
  12. Chang Y. Y, Eichel J., Cronan J. E. Jr 2000; Metabolic instability of Escherichia coli cyclopropane fatty acid synthase is due to RpoH-dependent proteolysis. J Bacteriol 182:4288–4294
    [Google Scholar]
  13. 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
    [Google Scholar]
  14. Cox J. S, Chen B, McNeil M., Jacobs W. R. Jr 1999; Complex lipid determinates tissue-specific replication of Mycobacterium tuberculosis in mice. Nature 402:79–83
    [Google Scholar]
  15. Cronan J. E. 2002; Phospholipid modifications in bacteria. Curr Opin Microbiol 5:202–205
    [Google Scholar]
  16. Daffé M., Draper P. 1998; The envelope layers of mycobacteria with reference to their pathogenicity. Adv Microbiol Phys 39:131–203
    [Google Scholar]
  17. Davidson L. A, Draper P., Minnikin D. E. 1982; Studies on the mycolic acids from the walls of Mycobacterium microti . J Gen Microbiol 128:823–828
    [Google Scholar]
  18. Dubnau E, Chan J, Raynaud C, Mohan V. P, Lanéelle M. A, Yu K, Quémard A, Smith I., Daffé M. 2000; Oxygenated mycolic acids are necessary for virulence of Mycobacterium tuberculosis in mice. Mol Microbiol 36:630–637
    [Google Scholar]
  19. George K. M, Yuan Y, Sherman D. R., Barry C. E.III. 1995; The biosynthesis of cyclopropanated mycolic acids in Mycobacterium tuberculosis . Identification and functional analysis of CMAS-2. J Biol Chem 270:27292–27298
    [Google Scholar]
  20. Gilleron M, Ronet C, Mempel M, Monsarrat B, Gachelin G., Puzo G. 2001; Acylation state of the phosphatidylinositol mannosides from Mycobacterium bovis Calmette Guérin and ability to induce granuloma and recruit natural killer T cells. J Biol Chem 276:34896–34904
    [Google Scholar]
  21. Glickman M. S, Cox J. S., Jacobs W. R. Jr 2000; A novel mycolic acid cyclopropane synthetase is required for cording, persistence, and virulence of Mycobacterium tuberculosis . Mol Cell 5:717–727
    [Google Scholar]
  22. Glickman M. S, Cahill S. M., Jacobs W. R. Jr 2001; The Mycobacterium tuberculosis cmaA2 gene encodes a mycolic acid trans -cyclopropane synthetase. J Biol Chem 276:2228–2233
    [Google Scholar]
  23. Gurcha S. S, Baulard A. R, Kremer L. 7 other authors 2002; Ppm1, a novel polyprenol monophosphomannose synthase from Mycobacterium tuberculosis . Biochem J 365:441–450
    [Google Scholar]
  24. Hoppe H. C, De Wet B. J. M, Cywes C, Daffé M., Ehlers M. R. W. 1997; Identification of phosphatidylinositol mannoside as a mycobacterial adhesin mediating both direct and opsonic binding to nonphagocytic mammalian cells. Infect Immun 65:3896–3905
    [Google Scholar]
  25. Huygen K, Lozes E, Gilles B. 10 other authors 1994; Mapping of TH1 helper T-cell epitopes on major secreted mycobacterial antigen 85A in mice infected with live Mycobacterium bovis BCG. Infect Immun 62:363–370
    [Google Scholar]
  26. Kikuchi S, Rainwater D. L., Kolattukudy P. E. 1992; Purification and characterization of an unusually large fatty acid synthase from Mycobacterium tuberculosis var. bovis BCG. Arch Biochem Biophys 295:318–326
    [Google Scholar]
  27. Kremer L, Baulard A, Estaquier J, Content J, Capron A., Locht C. 1995; Analysis of the Mycobacterium tuberculosis 85A Antigen promoter region. J Bacteriol 177:642–653
    [Google Scholar]
  28. Kremer L, Baulard A. R., Besra G. S. 2000; Genetics of mycolic acid biosynthesis. In Molecular Genetics of Mycobacteria pp 173–190 Edited by Hatfull G. F., Jacobs W. R. Jr Washington, DC: American Society for Microbiology;
    [Google Scholar]
  29. Kremer L, Dover L, Carrère S. 7 other authors 2002a; Mycolic acid biosynthesis and enzymatic characterization of the beta-ketoacyl-ACP synthase A-condensing enzyme from Mycobacterium tuberculosis. Biochem J 364:423–430
    [Google Scholar]
  30. Kremer L, Maughan W. N, Wilson R. A, Dover L. G., Besra G. S. 2002b; The M. tuberculosis antigen 85 complex and mycolyltransferase activity. Lett Appl Microbiol 34:233–237
    [Google Scholar]
  31. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  32. Levy-Frebault V, Daffé M, Restrepo E, Grimont F, Grimont P. A., David H. L. 1986; Differentiation of Mycobacterium thermoresistibile from Mycobacterium phlei and other rapidly growing mycobacteria. Ann Inst Pasteur Microbiol 137A:143–151
    [Google Scholar]
  33. Magnuson K, Jackowski S, Rock C. O., Cronan J. E. Jr 1993; Regulation of fatty acid biosynthesis in Escherichia coli . Microbiol Rev 57:522–542
    [Google Scholar]
  34. Marr A. G., Ingrahm J. L. 1962; Effect of temperature on the composition of fatty acids in Escherichia coli . J Bacteriol 84:1260–1267
    [Google Scholar]
  35. Minnikin D. E. 1982; Lipids: complex lipids, their chemistry, biosynthesis and roles. In The Biology of the Mycobacteria pp 95–184 Edited by Ratledge C., Stanford J. London: Academic Press;
    [Google Scholar]
  36. Peterson D. O., Bloch K. 1977; Mycobacterium smegmatis fatty acid synthetase: long chain transacylase chain length specificity. J Biol Chem 252:5735–5739
    [Google Scholar]
  37. Schaeffer M. L, Agnihotri G, Volker C, Kallender H, Brennan P. J., Lonsdale J. T. 2001; Purification and biochemical characterization of the Mycobacterium tuberculosis beta-ketoacyl-acyl carrier protein synthases. KasA and KasB. J Biol Chem 276:47029–47037
    [Google Scholar]
  38. Silvius J. R., McElhaney R. N. 1979; Effects of phospholipid acyl chain structure on thermotropic phase properties. 2. Phosphatidylcholines with unsaturated or cyclopropane acyl chains. Chem Phys Lipids 25:125–134
    [Google Scholar]
  39. Toriyama S, Yano I, Masui M, Kusunose M., Kusunose E. 1978; Separation of C50–60 and C70–80 mycolic acid molecular species and their changes by growth temperatures in Mycobacterium phlei . FEBS Lett 95:111–115
    [Google Scholar]
  40. Toriyama S, Yano I, Masui M, Kusunose E, Kusunose M., Akimori N. 1980; Regulation of cell wall mycolic acid biosynthesis in acid-fast bacteria. I. Temperature-induced changes in mycolic acid molecular species and related compounds in Mycobacterium phlei . J Biochem 88:211–221
    [Google Scholar]
  41. Wiker H. G., Harboe M. 1992; The antigen 85 complex: a major secretion product of Mycobacterium tuberculosis . Microbiol Rev 56:648–661
    [Google Scholar]
  42. Yuan Y, Zhu Y, Crane D. D., Barry C. E.III. 1998; The effect of oxygenated mycolic acid composition on cell wall function and macrophage growth in Mycobacterium tuberculosis . Mol Microbiol 29:1449–1458
    [Google Scholar]
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