1887

Microbiology

Volume 145, Issue 11

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

The EMBL accession number for the sequence reported in this paper is AJ132503.

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.

  • Received:
  • Accepted:
  • Revised:
  • 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
© Microbiology Society
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-145-11-3059
1999-11-01
2020-09-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/145/11/1453059a.html?itemId=/content/journal/micro/10.1099/00221287-145-11-3059&mimeType=html&fmt=ahah

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. Microbiology141:299–309[CrossRef]
     [Google Scholar]
  2. Altschul S. F., Gish W., Miller W., Myers W., Lipman D. J.. 1990; Basic local alignment search tool. J Mol Biol251:403–410
     [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 Res18:3642–3642[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 J8:2727–2736
     [Google Scholar]
  5. Donadio S., Katz L.. 1992; Organization of the enzymatic domain in multifunctional polyketide synthase involved in erythromycin formation in Saccharopolyspora erythraea. Gene111:51–60[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. Science252:675–679[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 USA92:3338–3342[CrossRef]
     [Google Scholar]
  8. Fytlovtich S., Nathan P. D., Zafriri D., Rosenberg E.. 1982; Amino acid precursors of Myxococcus xanthus antibiotic TA. J Antibiot35:1525–1530
     [Google Scholar]
  9. Hopwood D. A., Sherman D. H.. 1990; Molecular genetics of polyketides and its comparison to fatty acid biosynthesis. Annu Rev Genet24:37–66[CrossRef]
     [Google Scholar]
  10. Kashefi A., Hartzell P. L.. 1995; Genetic suppression and phenotypic masking of a Myxococcus xanthus frzF-defect. Mol Microbiol15:483–494[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 Biophys283:523–529[CrossRef]
     [Google Scholar]
  12. Kroos L., Kuspa A., Kaiser D.. 1986; A global analysis of developmentally regulated genes in Myxococcus xanthus. Dev Biol117:252–266[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. Nature325:818–821[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 Periodontol16:621–624[CrossRef]
     [Google Scholar]
  15. Norrander J., Kempe T., Messing J.. 1983; Construction of improved M13 vector. Gene26:101–106[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. Microbiology144:1641–1647[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 Biol286:465–474[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. Gene228:147–153[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 Lett170:221–227[CrossRef]
     [Google Scholar]
  20. Pearson W. R.. 1990; Rapid and sensitive sequence comparison with fastp and fasta. Methods Enzymol183:63–98
     [Google Scholar]
  21. Reichenbach H., Hofle G.. 1993; Production of bioactive secondary metabolites. In Myxobacteria II pp.347–398Edited by Dworkin M., Kaiser D.. Washington, DC: American Society for Microbiology.;
     [Google Scholar]
  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 Genet257:641–648[CrossRef]
     [Google Scholar]
  23. Rosenberg E., Varon M.. 1984; Antibiotics and lytic enzymes. In Myxobacteria, Development and Cell Interaction pp.109–125Edited by Rosenberg E.. New York: Springer;
     [Google Scholar]
  24. Rosenberg E., Vaks B., Zuckerberg A.. 1973; Bactericidal action of an antibiotic produced by Myxococcus xanthus. . Antimicrob Agents Chemother4:507–513[CrossRef]
     [Google Scholar]
  25. Rosenberg E., Fytlovitch S., Carmeli S., Kashman Y.. 1982; Chemical properties of Myxococcus xanthus antibotic TA. J Antibiot35:788–793[CrossRef]
     [Google Scholar]
  26. Rosenberg E., Porter J. M., Nathan P. N., Manor A., Varon M.. 1984; Antibiotic TA: an adherent antibiotic. Bio/Technology2:796–799[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 Acta1132:329–31[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;
     [Google Scholar]
  29. Sanger F., Nicklen S., Coulson A. R.. 1977; DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA74:5463–5467[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 Bacteriol177:3673–3679
     [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. Gene130:65–71[CrossRef]
     [Google Scholar]
  32. Sedgwick S. G., Morgan B. A.. 1994; Locating, DNA seqencing and disrupting yeast genes using tagged Tn1000. Methods Mol Genet3:131–140
     [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 J8:2717–2725
     [Google Scholar]
  34. Shimkets L. J.. 1990; Social and developmental biology of myxobacteria. Microbiol Rev54:473–501
     [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 Chemother36:2322–2327[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 Commun1982:1340–1342
     [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 Chem267:6807–6814
     [Google Scholar]
  38. Varon M., Rosenberg E.. 1996; Transcriptional regulation of genes required for antibiotic TA synthesis in Myxococcus xanthus. FEMS Microbiol Lett136:203–208[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 Chemother36:2316–2321[CrossRef]
     [Google Scholar]
  40. Varon M., Paitan Y., Rosenberg E.. 1997; Trans-acting regulation of antibiotic TA genes in Myxococcus xanthus. FEMS Microbiology Lett155:141–146[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 USA88:10783–10787[CrossRef]
     [Google Scholar]
  42. Zafriri D., Rosenberg E., Mirelman D.. 1981; Mode of action of Myxococcus xanthus antibiotic TA. Antimicrob Agents Chemother19:349–351[CrossRef]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-145-11-3059
Loading
Genetic and functional analysis of genes required for the post-modification of the polyketide antibiotic TA of Myxococcus xanthusThe EMBL accession number for the sequence reported in this paper is AJ132503.
Microbiology 145, 3059 (1999); https://doi.org/10.1099/00221287-145-11-3059
/content/journal/micro/10.1099/00221287-145-11-3059
/content/journal/micro/10.1099/00221287-145-11-3059
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