1887

Microbiology

Volume 145, Issue 11

Genetic and functional analysis of genes required for the post-modification of the polyketide antibiotic TA of Free

Abstract

The antibiotic TA of is a complex macrocyclic polyketide, produced through successive condensations of acetate by a type I PKS (polyketide synthase) mechanism. The genes encoding TA biosynthesis are clustered on a 36 kb DNA fragment, which has been cloned and analysed. The chemical structure of TA and the mechanism by which it is synthesized indicate the need for several post-modification steps, which are introduced into the carbon chain of the polyketide to form the final bioactive molecule. These include the addition of several carbon atoms originating from acetate carbonyl, three C-methylations, O-methylation and a specific hydroxylation. This paper reports the analysis of five genes which are involved in the post-modification of TA. Their functional analysis, by specific gene disruption, suggests that they may be essential for the production of the active antibiotic. The characteristics and organization of the genes suggest that they may be involved in the addition of the carbon atoms which arise from acetate.

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  • Accepted:
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  • Published Online:
Keyword(s): β-ketoacyl ACP synthase III , ACP, acyl carrier protein , acyl carrier protein , antibiotic post-modification , HMG, hydroxymethylglutaryl , KAS, β-ketoacyl:ACP synthase , Myxococcus xanthus , PKS, polyketide synthase and polyketide synthase
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1999-11-01
2019-12-15
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References

  1. Albertini, A. M., Caramori, T., Scoffone, F., Scotti, C. & Galizzi, A. ( 1995; ). Sequence around the 159° region of the Bacillus subtilis genome: the pksX locus spans 33·6 kb. Microbiology 141, 299-309.[CrossRef]
    [Google Scholar]
  2. Altschul, S. F., Gish, W., Miller, W., Myers, W. & Lipman, D. J. ( 1990; ). Basic local alignment search tool. J Mol Biol 251, 403-410.
     [Google Scholar]
  3. Ayte, J., Gil-Gomez, G. & Hegardt, F. G. ( 1990; ). Nucleotide sequence of a rat liver cDNA encoding the cytosolic 3-hydroxy-3-methylglutaryl coenzyme A synthase. Nucleic Acids Res 18, 3642-3642.[CrossRef]
    [Google Scholar]
  4. Bibb, M. J., Biro, S., Motamedi, H., Collins, J. F. & Hutchinson, C. R. ( 1989; ). Analysis of the nucleotide sequence of the Streptomyces glaucescens tcmI genes provides key information about the enzymology of polyketide antibiotic biosynthesis. EMBO J 8, 2727-2736.
     [Google Scholar]
  5. Donadio, S. & Katz, L. ( 1992; ). Organization of the enzymatic domain in multifunctional polyketide synthase involved in erythromycin formation in Saccharopolyspora erythraea. Gene 111, 51-60.[CrossRef]
    [Google Scholar]
  6. Donadio, S., Staver, M. J., McAlpine, J. B., Swanson, S. J. & Katz, L. ( 1991; ). Modular organization of genes required for complex polyketide biosynthesis. Science 252, 675-679.[CrossRef]
    [Google Scholar]
  7. Durbin, M. L., Learn, G. H.Jr, Huttley, G. A. & Clegg, M. T. ( 1995; ). Evolution of the chalcone synthase gene family in the genus Ipomoea. Proc Natl Acad Sci USA 92, 3338-3342.[CrossRef]
    [Google Scholar]
  8. Fytlovtich, S., Nathan, P. D., Zafriri, D. & Rosenberg, E. ( 1982; ). Amino acid precursors of Myxococcus xanthus antibiotic TA. J Antibiot 35, 1525-1530.
     [Google Scholar]
  9. Hopwood, D. A. & Sherman, D. H. ( 1990; ). Molecular genetics of polyketides and its comparison to fatty acid biosynthesis. Annu Rev Genet 24, 37-66.[CrossRef]
    [Google Scholar]
  10. Kashefi, A. & Hartzell, P. L. ( 1995; ). Genetic suppression and phenotypic masking of a Myxococcus xanthus frzF-defect. Mol Microbiol 15, 483-494.[CrossRef]
    [Google Scholar]
  11. Kattar-Cooley, P. A., Wang, H. H., Mende-Mueller, L. M. & Miziorko, H. M. ( 1990; ). Avian liver 3-hydroxy-3-methylglutaryl-CoA synthase: distinct genes encode the cholesterogenic and ketogenic isozymes. Arch Biochem Biophys 283, 523-529.[CrossRef]
    [Google Scholar]
  12. Kroos, L., Kuspa, A. & Kaiser, D. ( 1986; ). A global analysis of developmentally regulated genes in Myxococcus xanthus. Dev Biol 117, 252-266.[CrossRef]
    [Google Scholar]
  13. Malpartida, F., Hallam, S. E., Kieser, H. M. & 9 other authors ( 1987; ). Homology between Streptomyces genes coding for synthesis of different polyketides used to clone antibiotic biosynthesis genes. Nature 325, 818–821.[CrossRef]
    [Google Scholar]
  14. Manor, A., Eli, I., Varon, M., Judes, H. & Rosenberg, E. ( 1989; ). Effect of adhesive antibiotic TA on plaque and gingivitis in man. J Clin Periodontol 16, 621-624.[CrossRef]
    [Google Scholar]
  15. Norrander, J., Kempe, T. & Messing, J. ( 1983; ). Construction of improved M13 vector. Gene 26, 101-106.[CrossRef]
    [Google Scholar]
  16. Paitan, Y., Boulton, N., Ron, E. Z., Rosenberg, E. & Orr, E. ( 1998; ). Molecular analysis of the gyrB gene from Myxococcus xanthus. Microbiology 144, 1641-1647.[CrossRef]
    [Google Scholar]
  17. Paitan, Y., Alon, G., Orr, E., Ron, E. Z. & Rosenberg, E. ( 1999a; ). The first gene in the biosynthesis of the polyketide antibiotic TA of Myxococcus xanthus codes for a unique PKS module coupled to a peptide synthetase. J Mol Biol 286, 465-474.[CrossRef]
    [Google Scholar]
  18. Paitan, Y., Orr, E., Ron, E. Z. & Rosenberg, E. ( 1999b; ). Cloning and characterization of a Myxococcus xanthus cytochrome P-450 hydroxylase required for biosynthesis of the polyketide antibiotic TA. Gene 228, 147-153.[CrossRef]
    [Google Scholar]
  19. Paitan, Y., Orr, E., Ron, E. Z. & Rosenberg, E. ( 1999c; ). A NusG-like transcription anti-terminator is involved in the biosynthesis of the polyketide antibiotic TA of Myxococcus xanthus. FEMS Microbiol Lett 170, 221-227.[CrossRef]
    [Google Scholar]
  20. Pearson, W. R. ( 1990; ). Rapid and sensitive sequence comparison with fastp and fasta. Methods Enzymol 183, 63-98.
     [Google Scholar]
  21. Reichenbach, H. & Hofle, G. ( 1993; ). Production of bioactive secondary metabolites. In Myxobacteria II, pp. 347-398. Edited by M. Dworkin & D. Kaiser. Washington, DC: American Society for Microbiology.
  22. Ritsema, T., Gehring, A. M., Stuitje, A. R. & 7 other authors ( 1998; ). Functional analysis of an interspecies chimera of acyl carrier proteins indicate a specialized domain for protein recognition. Mol Gen Genet 257, 641–648.[CrossRef]
    [Google Scholar]
  23. Rosenberg, E. & Varon, M. ( 1984; ). Antibiotics and lytic enzymes. In Myxobacteria, Development and Cell Interaction, pp. 109-125. Edited by E. Rosenberg. New York: Springer.
  24. Rosenberg, E., Vaks, B. & Zuckerberg, A. ( 1973; ). Bactericidal action of an antibiotic produced by Myxococcus xanthus. Antimicrob Agents Chemother 4, 507-513.[CrossRef]
    [Google Scholar]
  25. Rosenberg, E., Fytlovitch, S., Carmeli, S. & Kashman, Y. ( 1982; ). Chemical properties of Myxococcus xanthus antibotic TA. J Antibiot 35, 788-793.[CrossRef]
    [Google Scholar]
  26. Rosenberg, E., Porter, J. M., Nathan, P. N., Manor, A. & Varon, M. ( 1984; ). Antibiotic TA: an adherent antibiotic. Bio/Technology 2, 796-799.[CrossRef]
    [Google Scholar]
  27. Russ, A. P., Ruzicka, V., Maerz, W., Appelhans, H. & Gross, W. ( 1992; ). Amplification and direct sequencing of a cDNA encoding human cytosolic 3-hydroxy-3-methylglutaryl-coenzyme A synthase. Biochim Biophys Acta 1132, 329-31.[CrossRef]
    [Google Scholar]
  28. Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  29. Sanger, F., Nicklen, S. & Coulson, A. R. ( 1977; ). DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74, 5463-5467.[CrossRef]
    [Google Scholar]
  30. Schupp, T., Toupet, C., Cluzel, B., Neff, S., Hill, S., Back, J. J. & Ligon, J. M. ( 1995; ). A Sorangium cellulosum (Myxobacterium) gene cluster for the biosynthesis of the macrolide antibiotic Soraphen A: cloning, characterization, and homology to polyketide synthase genes from Actinomycetes. J Bacteriol 177, 3673-3679.
     [Google Scholar]
  31. Scotti, C., Piatti, M., Cuzzoni, A., Perani, P., Tognoni, A., Grandi, G., Galizzi, A. & Albertini, A. M. ( 1993; ). A Bacillus subtilis large ORF coding for a polypeptide highly similar to polyketide synthases. Gene 130, 65-71.[CrossRef]
    [Google Scholar]
  32. Sedgwick, S. G. & Morgan, B. A. ( 1994; ). Locating, DNA seqencing and disrupting yeast genes using tagged Tn1000. Methods Mol Genet 3, 131-140.
     [Google Scholar]
  33. Sherman, D. H., Malpartida, F., Bibb, M. J., Kieser, H. M. & Hopwood, D. A. ( 1989; ). Structure and deduced function of the granaticin-producing polyketide synthase gene cluster of Streptomyces violaceoruber Tü22. EMBO J 8, 2717-2725.
     [Google Scholar]
  34. Shimkets, L. J. ( 1990; ). Social and developmental biology of myxobacteria. Microbiol Rev 54, 473-501.
     [Google Scholar]
  35. Tolchinsky, S., Fuchs, N., Varon, M. & Rosenberg, E. ( 1992; ). Use of Tn5lac to study expression of genes required for antibiotic TA production. Antimicrob Agents Chemother 36, 2322-2327.[CrossRef]
    [Google Scholar]
  36. Trowitzsch, W., Wray, V., Gerth, K. & Hofle, G. ( 1982; ). Structure of Myxovirescin A, a new macrocyclic antibiotic from gliding bacteria. J Chem Soc Commun 1982, 1340-1342.
     [Google Scholar]
  37. Tsay, J. T., Oh, W., Larson, T. J., Jackowski, S. & Rock, C. O. ( 1992; ). Isolation and characterization of the beta-ketoacyl-acyl carrier protein synthase III gene (fabH) from Escherichia coli K-12. J Biol Chem 267, 6807-6814.
     [Google Scholar]
  38. Varon, M. & Rosenberg, E. ( 1996; ). Transcriptional regulation of genes required for antibiotic TA synthesis in Myxococcus xanthus. FEMS Microbiol Lett 136, 203-208.[CrossRef]
    [Google Scholar]
  39. Varon, M., Fuchs, N., Monosov, M., Tolchinsky, S. & Rosenberg, E. ( 1992; ). Mutation and mapping of genes involved in antibiotic TA production in Myxococcus xanthus. Antimicrob Agents Chemother 36, 2316-2321.[CrossRef]
    [Google Scholar]
  40. Varon, M., Paitan, Y. & Rosenberg, E. ( 1997; ). Trans-acting regulation of antibiotic TA genes in Myxococcus xanthus. FEMS Microbiology Lett 155, 141-146.[CrossRef]
    [Google Scholar]
  41. Wang, S. L. & Liu, X. Q. ( 1991; ). The plastid genome of Cryptomonas phi encodes an hsp70-like protein, a histone-like protein, and an acyl carrier protein. Proc Natl Acad Sci USA 88, 10783-10787.[CrossRef]
    [Google Scholar]
  42. Zafriri, D., Rosenberg, E. & Mirelman, D. ( 1981; ). Mode of action of Myxococcus xanthus antibiotic TA. Antimicrob Agents Chemother 19, 349-351.[CrossRef]
    [Google Scholar]
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Genetic and functional analysis of genes required for the post-modification of the polyketide antibiotic TA of Myxococcus xanthus
Microbiology 145, 3059 (1999); https://doi.org/10.1099/00221287-145-11-3059
/content/journal/micro/10.1099/00221287-145-11-3059
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