1887

Abstract

The methyl-branched fatty acyl components of sulfolipid-I (SL-I), a major glycolipid of the human pathogen , are synthesized by the polyketide synthase Pks2. (), located downstream of , encodes a protein that belongs to a subfamily of acyltransferases associated with mycobacterial polyketide synthases [polyketide synthase-associated proteins (PAPs)]. The presence of a conserved acyltransferase motif (HXDXY) suggested a role for PapA1 in acylation of sulfated trehalose to form SL-I. Targeted deletion of the H37Rv resulted in loss of SL-I, demonstrating its role in mycobacterial sulfolipid biosynthesis. Furthermore, SL-I synthesis was restored in the mutant strain following complementation with , but not with mutant alleles of containing alterations of key residues in the acyltransferase motif, confirming that PapA1 was an acyltransferase. While other clusters are associated with a single PAP-encoding gene, it was demonstrated that another open reading frame, (), located 5.8 kb downstream of is also an acyltransferase gene involved in SL-I biosynthesis: deletion of abolished SL-I production. The absence of any partially acylated intermediates in either null mutant indicated that both PapA1 and PapA2 were required for all acylation steps of SL-I assembly.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.2006/003103-0
2007-02-01
2020-08-10
Loading full text...

Full text loading...

/deliver/fulltext/micro/153/2/513.html?itemId=/content/journal/micro/10.1099/mic.0.2006/003103-0&mimeType=html&fmt=ahah

References

  1. Bardarov S., Sambandamurthy V., Larsen M., Tufariello J., Chan J., Hatfull G., Jacobs W. R. Jr, Bardarov S. Jr, Pavelka M. S. Jr. 2002; Specialized transduction: an efficient method for generating marked and unmarked targeted gene disruptions in Mycobacterium tuberculosis , M.bovis BCG and M. smegmatis . Microbiology148:3007–3017
    [Google Scholar]
  2. Bergendahl V., Linne U., Marahiel M. A.. 2002; Mutational analysis of the C-domain in nonribosomal peptide synthesis. Eur J Biochem269:620–629[CrossRef]
    [Google Scholar]
  3. Brennan P. J., Nikaido H.. 1995; The envelope of mycobacteria. Annu Rev Biochem64:29–63[CrossRef]
    [Google Scholar]
  4. Cole S. T., Brosch R., Parkhill J. Garnier T., Churcher C. Harris D., Gordon S. V., Eiglmeier K., Gas S.. other authors 1998; Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature393:537–544[CrossRef]
    [Google Scholar]
  5. Converse S. E., Mougous J. D., Leavell M. D., Leary J. A., Bertozzi C. R., Cox J. S.. 2003; MmpL8 is required for sulfolipid-1 biosynthesis and Mycobacterium tuberculosis virulence. Proc Natl Acad Sci U S A100:6121–6126[CrossRef]
    [Google Scholar]
  6. Dobson G., Minnikin D. E., Minnikin S. M., Parlett M., Goodfellow M., Ridell M., Magnusson M.. 1985; Systematic analysis of complex mycobacterial lipids. In Chemical Methods in Bacterial Systematics pp237–265 Edited by Goodfellow M., Minnikin D. E.. London: Academic Press;
    [Google Scholar]
  7. Domenech P., Reed M. B., Dowd C. S., Manca C., Kaplan G., Barry C. E. 3rd. 2004; The role of MmpL8 in sulfatide biogenesis and virulence of Mycobacterium tuberculosis . J Biol Chem279:21257–21265[CrossRef]
    [Google Scholar]
  8. Garnier T., Eiglmeier K., Camus J. C., Medina N., Mansoor H., Prior M., Duthoy S., Grondin S., Lacroix C.. other authors 2003; The complete genome sequence of Mycobacterium bovis . Proc Natl Acad Sci U S A100:7877–7882[CrossRef]
    [Google Scholar]
  9. Goren M. B.. 1970a; Sulfolipid I of Mycobacterium tuberculosis , strain H37Rv. II. Structural studies. Biochim Biophys Acta210:127–138[CrossRef]
    [Google Scholar]
  10. Goren M. B.. 1970b; Sulfolipid I of Mycobacterium tuberculosis , strain H37Rv. I. Purification and properties. Biochim Biophys Acta210:116–126[CrossRef]
    [Google Scholar]
  11. Goren M. B., Brokl O., Das B. C., Lederer E.. 1971; Sulfolipid I of Mycobacterium tuberculosis , strain H37RV. Nature of the acyl substituents. Biochemistry10:72–81[CrossRef]
    [Google Scholar]
  12. Goren M. B., Brokl O., Das B. C.. 1976; Sulfatides of Mycobacterium tuberculosis : the structure of the principal sulfatide (SL-I. Biochemistry15:2728–2735[CrossRef]
    [Google Scholar]
  13. Jain M., Cox J. S.. 2005; Interaction between polyketide synthase and transporter suggests coupled synthesis and export of virulence lipid in M. tuberculosis . PLoS Pathog1:e2[CrossRef]
    [Google Scholar]
  14. Lewendon A., Murray I. A., Kleanthous C., Cullis P. M., Shaw W. V.. 1988; Substitutions in the active site of chloramphenicol acetyltransferase: role of a conserved aspartate. Biochemistry27:7385–7390[CrossRef]
    [Google Scholar]
  15. Middlebrook G., Coleman C. M., Schaefer W. B.. 1959; Sulfolipid from virulent tubercle bacilli. Proc Natl Acad Sci U S A45:1801–1804[CrossRef]
    [Google Scholar]
  16. Mougous J. D., Petzold C. J., Senaratne R. H., Lee D. H., Akey D. L., Lin F. L., Munchel S. E., Pratt M. R., Riley L. W.. other authors 2004; Identification, function and structure of the mycobacterial sulfotransferase that initiates sulfolipid-1 biosynthesis. Nat Struct Mol Biol11:721–729[CrossRef]
    [Google Scholar]
  17. Okamoto Y., Fujita Y., Naka T., Hirai M., Tomiyasu I., Yano I.. 2006; Mycobacterial sulfolipid shows a virulence by inhibiting cord factor induced granuloma formation and TNF-alpha release. Microb Pathog40:245–253[CrossRef]
    [Google Scholar]
  18. Onwueme K. C., Ferreras J. A., Buglino J., Lima C. D., Quadri L. E.. 2004; Mycobacterial polyketide-associated proteins are acyltransferases: proof of principle with Mycobacterium tuberculosis PapA5. Proc Natl Acad Sci U S A101:4608–4613[CrossRef]
    [Google Scholar]
  19. Pabst M. J., Gross J. M., Brozna J. P., Goren M. B.. 1988; Inhibition of macrophage priming by sulfatide from Mycobacterium tuberculosis . J Immunol140:634–640
    [Google Scholar]
  20. Puzo G.. 1990; The carbohydrate- and lipid-containing cell wall of mycobacteria, phenolic glycolipids: structure and immunological properties. Crit Rev Microbiol17:305–327[CrossRef]
    [Google Scholar]
  21. Rivera-Marrero C. A., Ritzenthaler J. D., Newburn S. A., Roman J., Cummings R. D.. 2002; Molecular cloning and expression of a novel glycolipid sulfotransferase in Mycobacterium tuberculosis . Microbiology148:783–792
    [Google Scholar]
  22. Rousseau C., Turner O. C., Rush E., Bordat Y., Sirakova T. D., Kolattukudy P. E., Ritter S., Orme I. M., Giquel B., Jackson M.. 2003; Sulfolipid deficiency does not affect the virulence of Mycobacterium tuberculosis H37Rv in mice and guinea pigs. Infect Immun71:4684–4690[CrossRef]
    [Google Scholar]
  23. Sambandamurthy V. K., Wang X., Chen B., Russell R. G., Derrick S., Collins F. M., Morris S. L., Jacobs W. R. Jr. 2002; A pantothenate auxotroph of Mycobacterium tuberculosis is highly attenuated and protects mice against tuberculosis. Nat Med8:1171–1174[CrossRef]
    [Google Scholar]
  24. Sirakova T. D., Thirumala A. K., Dubey V. S., Sprecher H., Kolattukudy P. E.. 2001; The Mycobacterium tuberculosis pks2 gene encodes the synthase for the hepta- and octamethyl-branched fatty acids required for sulfolipid synthesis. J Biol Chem276:16833–16839[CrossRef]
    [Google Scholar]
  25. Stover C. K., Fuerst T. R., Burlein J. E., Benson L. A., Bennett L. T., Bansal G. P., Young J. F., Lee M. H., other authors de la Cruz V. F.. 1991; New use of BCG for recombinant vaccines. Nature351:456–460[CrossRef]
    [Google Scholar]
  26. Trivedi O. A., Arora P., Vats A., Ansari M. Z., Tickoo R., Sridharan V., Mohanty D., Gokhale R. S.. 2005; Dissecting the mechanism and assembly of a complex virulence mycobacterial lipid. Mol Cell17:631–643[CrossRef]
    [Google Scholar]
  27. Zhang L., Goren M. B., Holzer T. J., Andersen B. R.. 1988; Effect of Mycobacterium tuberculosis -derived sulfolipid I on human phagocytic cells. Infect Immun56:2876–2883
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.2006/003103-0
Loading
/content/journal/micro/10.1099/mic.0.2006/003103-0
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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