Lipidomics reveals control of Mycobacterium tuberculosis virulence lipids via metabolic coupling

Mycobacterium tuberculosis synthesizes specific polyketide lipids that interact with the host and are required for virulence. Using a mass spectrometric approach to simultaneously monitor hundreds of lipids, we discovered that the size and abundance of two lipid virulence factors, phthiocerol dimycocerosate (PDIM) and sulfolipid-1 (SL-1), are controlled by the availability of a common precursor, methyl malonyl CoA (MMCoA). Consistent with this view, increased levels of MMCoA led to increased abundance and mass of both PDIM and SL-1. Furthermore, perturbation of MMCoA metabolism attenuated pathogen replication in mice. Importantly, we detected increased PDIM synthesis in bacteria growing within host tissues and in bacteria grown in culture on odd-chain fatty acids. Because M. tuberculosis catabolizes host lipids to grow during infection, we propose that growth of M. tuberculosis on fatty acids in vivo leads to increased flux of MMCoA through lipid biosynthetic pathways, resulting in increased virulence lipid synthesis. Our results suggest that the shift to host lipid catabolism during infection allows for increased virulence lipid anabolism by the bacterium.

[1]  Gilla Kaplan,et al.  Differential expression of iron-, carbon-, and oxygen-responsive mycobacterial genes in the lungs of chronically infected mice and tuberculosis patients , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Gilla Kaplan,et al.  The Role of MmpL8 in Sulfatide Biogenesis and Virulence of Mycobacterium tuberculosis* , 2004, Journal of Biological Chemistry.

[3]  P. Kolattukudy,et al.  Fatty acid biosynthesis in Mycobacterium tuberculosis var. bovis Bacillus Calmette-Guérin. Purification and characterization of a novel fatty acid synthase, mycocerosic acid synthase, which elongates n-fatty acyl-CoA with methylmalonyl-CoA. , 1985, The Journal of biological chemistry.

[4]  John Chan,et al.  Responses of Mycobacterium tuberculosis to Growth in the Mouse Lung , 2005, Infection and Immunity.

[5]  J. Cox,et al.  Interaction between Polyketide Synthase and Transporter Suggests Coupled Synthesis and Export of Virulence Lipid in M. tuberculosis , 2005, PLoS pathogens.

[6]  Mohd Zeeshan Ansari,et al.  Dissecting the mechanism and assembly of a complex virulence mycobacterial lipid. , 2005, Molecular cell.

[7]  G. Puzo,et al.  Lipoarabinomannans: from structure to biosynthesis. , 2003, Biochimie.

[8]  A. Azad,et al.  Targeted replacement of the mycocerosic acid synthase gene in Mycobacterium bovis BCG produces a mutant that lacks mycosides. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[9]  Brigitte Gicquel,et al.  Production of phthiocerol dimycocerosates protects Mycobacterium tuberculosis from the cidal activity of reactive nitrogen intermediates produced by macrophages and modulates the early immune response to infection , 2004, Cellular microbiology.

[10]  A. Azad,et al.  Gene Knockout Reveals a Novel Gene Cluster for the Synthesis of a Class of Cell Wall Lipids Unique to Pathogenic Mycobacteria* , 1997, The Journal of Biological Chemistry.

[11]  William R. Jacobs,et al.  Complex lipid determines tissue-specific replication of Mycobacterium tuberculosis in mice , 1999, Nature.

[12]  H. Sprecher,et al.  The Mycobacterium tuberculosis pks2 Gene Encodes the Synthase for the Hepta- and Octamethyl-branched Fatty Acids Required for Sulfolipid Synthesis* , 2001, The Journal of Biological Chemistry.

[13]  Pierre Baldi,et al.  Structure-based inhibitor design of AccD5, an essential acyl-CoA carboxylase carboxyltransferase domain of Mycobacterium tuberculosis. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[14]  J. Graham,et al.  Identification of Mycobacterium tuberculosis RNAs synthesized in response to phagocytosis by human macrophages by selective capture of transcribed sequences (SCOTS). , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[15]  B. Gicquel,et al.  Identification of a virulence gene cluster of Mycobacterium tuberculosis by signature‐tagged transposon mutagenesis , 1999, Molecular microbiology.

[16]  E. Muñoz-Elías,et al.  Role of the methylcitrate cycle in Mycobacterium tuberculosis metabolism, intracellular growth, and virulence , 2006, Molecular microbiology.

[17]  D. Russell,et al.  The interaction between Mycobacterium and the macrophage analyzed by two‐dimensional polyacrylamide gel electrophoresis , 1997, Electrophoresis.

[18]  W. Segal,et al.  BIOCHEMICAL DIFFERENTIATION OF MYCOBACTERIUM TUBERCULOSIS GROWN IN VIVO AND IN VITRO , 1956, Journal of bacteriology.

[19]  H. Desaire,et al.  Combined partial acid hydrolysis and electrospray ionization-mass spectrometry for the structural determination of oligosaccharides. , 2000, Analytical chemistry.

[20]  M. Reed,et al.  A glycolipid of hypervirulent tuberculosis strains that inhibits the innate immune response , 2004, Nature.

[21]  E. Muñoz-Elías,et al.  Mycobacterium tuberculosis isocitrate lyases 1 and 2 are jointly required for in vivo growth and virulence , 2005, Nature Medicine.

[22]  Spencer J. Williams,et al.  Discovery of sulfated metabolites in mycobacteria with a genetic and mass spectrometric approach , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[23]  Rajesh S. Gokhale,et al.  Enzymic activation and transfer of fatty acids as acyl-adenylates in mycobacteria , 2004, Nature.

[24]  H. Nikaido,et al.  The envelope of mycobacteria. , 1995, Annual review of biochemistry.

[25]  R. Tibshirani,et al.  Significance analysis of microarrays applied to the ionizing radiation response , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[26]  James C. Sacchettini,et al.  Persistence of Mycobacterium tuberculosis in macrophages and mice requires the glyoxylate shunt enzyme isocitrate lyase , 2000, Nature.

[27]  M. Goren.  Sulfolipid I of Mycobacterium tuberculosis, strain H37Rv. II. Structural studies. , 1970, Biochimica et biophysica acta.

[28]  Yang Liu,et al.  Transcriptional Adaptation of Mycobacterium tuberculosis within Macrophages , 2003, The Journal of experimental medicine.

[29]  Carolyn R Bertozzi,et al.  MmpL8 is required for sulfolipid-1 biosynthesis and Mycobacterium tuberculosis virulence , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[30]  E. Muñoz-Elías,et al.  Carbon metabolism of intracellular bacteria , 2006, Cellular microbiology.

[31]  Gary K. Schoolnik,et al.  ideR, an Essential Gene in Mycobacterium tuberculosis: Role of IdeR in Iron-Dependent Gene Expression, Iron Metabolism, and Oxidative Stress Response , 2002, Infection and Immunity.

[32]  A. Azad,et al.  Biochemistry and molecular genetics of cell‐wall lipid biosynthesis in mycobacteria , 1997, Molecular microbiology.

[33]  David G. Russell,et al.  Characterization of Activity and Expression of Isocitrate Lyase in Mycobacterium avium andMycobacterium tuberculosis , 1999, Journal of bacteriology.

[34]  W. J. Dyer,et al.  A rapid method of total lipid extraction and purification. , 1959, Canadian journal of biochemistry and physiology.