1887

Abstract

Amplification of sequences from ISP5230 genomic DNA using PCR with primers based on conserved prokaryotic sequences gave two main products. One matched , a locus previously identified in The second closely resembled the conserved sequence consensus and hybridized with a 38 kb I fragment of ISP5230 genomic DNA. Cloning and sequence analysis of the 38 kb fragment detected three ORFs, and their deduced amino acid sequences were used in BLAST searches of the GenBank database. The ORF1 product was similar to PabB in other bacteria and to the PabB domain encoded by . The ORF2 product resembled PabA of other bacteria. ORF3 was incomplete; its deduced partial amino acid sequence placed it in the MocR group of GntR-type transcriptional regulators. Introducing vectors containing the 38 kb I fragment of DNA into and mutants of , or into the mutant JG10, enhanced sulfanilamide resistance in the host strains. The increased resistance was attributed to expression of the pair of discrete translationally coupled -aminobenzoic acid biosynthesis genes (designated /) cloned in the 38 kb fragment. These represent a second set of genes encoding 4-amino-4-deoxychorismate synthase in ISP5230. In contrast to the fused set previously isolated from this species, they do not participate in chloramphenicol biosynthesis, but like they can be disrupted without affecting growth on minimal medium. The gene disruption results suggest that may have a third set of genes encoding PABA synthase.

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2001-08-01
2021-07-28
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References

  1. Aidoo D. A. 1989; Approaches to the cloning of genes for chloramphenicol biosynthesis in Streptomyces venezuelae ISP5230. PhD thesis Dalhousie University; Halifax, NS, Canada:
    [Google Scholar]
  2. Aidoo D. A., Barrett K., Vining L. C. 1990; Plasmid transformation of Streptomyces venezuelae : modified procedures used to introduce the genes for p -aminobenzoate synthetase. J Gen Microbiol 136:657–662 [CrossRef]
    [Google Scholar]
  3. Altschul S. F., Madden T. L., Schaffer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J. 1997; Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402 [CrossRef]
    [Google Scholar]
  4. Brown M. P., Aidoo K. A., Vining L. C. 1996; A role for pabAB , a p -aminobenzoate synthase gene of Streptomyces venezuelae ISP5230, in chloramphenicol biosynthesis. Microbiology 142:1345–1355 [CrossRef]
    [Google Scholar]
  5. Chang Z. 1999; Genes for cysteine biosynthesis and metabolism in Streptomyces venezuelae ISP5230: cloning, sequencing, functional analysis and relevance to chloramphenicol biosynthesis. PhD thesis Dalhousie University; Halifax, NS, Canada:
    [Google Scholar]
  6. Chong P. P., Podmore S. M., Kieser H. M., Redenbach M., Turgay K., Marahiel M., Hopwood D. A., Smith C. P. 1998; Physical identification of a chromosomal locus encoding biosynthetic genes for the lipopeptide calcium-dependent antibiotic (CDA) of Streptomyces coelicolor A3(2). Microbiology 144:193–199 [CrossRef]
    [Google Scholar]
  7. Clowes R. C., Hayes W. 1968 Experiments in Microbial Genetics. New York: Wiley;
    [Google Scholar]
  8. Devereux J., Haeberli P., Smithies O. 1984; A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395 [CrossRef]
    [Google Scholar]
  9. Doull J. L., Vats S., Chaliciopoulos M., Stuttard C., Wong K., Vining L. C. 1986; Conjugational fertility and location of chloramphenicol biosynthesis genes on the chromosomal linkage map of Streptomyces venezuelae. J Gen Microbiol 132:1327–1338 [CrossRef]
    [Google Scholar]
  10. Edman J. C., Goldstein A. L., Erbe J. G. 1993; Para-aminobenzoate synthase gene of Saccharomyces cerevisiae encodes a bifunctional enzyme. Yeast 9:669–675 [CrossRef]
    [Google Scholar]
  11. Flett F., Mersinias V., Smith C. P. 1997; High-efficiency intergeneric conjugal transfer of plasmid DNA from Escherichia coli to methyl DNA-restricting streptomycetes. FEMS Microbiol Lett 155:223–229 [CrossRef]
    [Google Scholar]
  12. Gil J. A., Hopwood D. A. 1983; Cloning and expression of a p- aminobenzoic acid synthase gene of the candicidin-producing Streptomyces griseus . Gene 25:119–132 [CrossRef]
    [Google Scholar]
  13. Goncharoff P., Nichols B. P. 1984; Nucleotide sequence of Escherichia coli pabB indicates a common evolutionary origin of p- aminobenzoate synthetase and anthranilate synthetase. J Bacteriol 159:57–62
    [Google Scholar]
  14. Green J. M., Merkel W. K., Nichols B. P. 1992; Characterization and sequence of Escherichia coli pabC , the gene encoding aminodeoxychorismate lyase, a pyridoxal phosphate-containing enzyme. J Bacteriol 174:5317–5323
    [Google Scholar]
  15. Halpern T. 1997; Role of p-aminophenylalanine in the biosynthesis of chloramphenicol. BSc Honours thesis Dalhousie University; Halifax, NS, Canada:
    [Google Scholar]
  16. Henikoff S. 1984; Unidirectional deletion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359 [CrossRef]
    [Google Scholar]
  17. Higgins D. G., Thompson J. D., Gibson T. J. 1996; Using clustal for multiple sequence alignments. Methods Enzymol 266:383–402
    [Google Scholar]
  18. Hopwood D. A. 1967; Genetic analysis and genome structure in Streptomyces coelicolor . Bacteriol Rev 31:373–403
    [Google Scholar]
  19. Hopwood D. A., Bibb M. J., Chater K. F., Kieser T., Bruton C. J., Kieser H. M., Lydiate D. J., Smith C. P., Ward J. M., Schrempf H. 1985 Genetic Manipulation of Streptomyces: a Laboratory Manual Norwich: John Innes Foundation;
    [Google Scholar]
  20. Jones A., Vining L. C. 1976; Biosynthesis of chloramphenicol in Streptomyces sp. 3022a. Identification of p -amino-l-phenylalanine as a product from the action of arylamine synthetase on chorismic acid. Can J Microbiol 22:237–244 [CrossRef]
    [Google Scholar]
  21. Larson J. L., Hershberger C. L. 1986; The minimal replicon of a streptomycete plasmid produces an ultrahigh level of plasmid DNA. Plasmid 15:199–209 [CrossRef]
    [Google Scholar]
  22. Lin C., Paradkar A. S., Vining L. C. 1998; Regulation of an anthranilate synthase gene in Streptomyces venezuelae by a trp attenuator. Microbiology 144:1971–1980 [CrossRef]
    [Google Scholar]
  23. MacNeil D. J., Gewain K. M., Rudy C. L., Dezeny G., Gibbons P. H., MacNeil T. 1992; Analysis of Streptomyces avermitilis genes required for avermectin biosynthesis utilizing a novel integration vector. Gene 111:61–68 [CrossRef]
    [Google Scholar]
  24. Malik V. S., Vining L. C. 1970; Metabolism of chloramphenicol by the producing organism. Can J Microbiol 16:173–179 [CrossRef]
    [Google Scholar]
  25. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  26. Mazodier P., Petter R., Thompson C. 1989; Intergeneric conjugation between Escherichia coli and Streptomyces species. J Bacteriol 171:3583–3585
    [Google Scholar]
  27. Munro M. H., Taniguchi G. M., Rinehart K. L., Gottlieb D. 1975; A cmr study of the biosynthesis of chloramphenicol. Tetrahedron Lett2659–2662
    [Google Scholar]
  28. Paradkar A. S., Jensen S. E. 1995; Functional analysis of the gene encoding the clavaminate synthase 2 isoenzyme involved in clavulanic acid biosynthesis in Streptomyces clavuligerus . J Bacteriol 177:1307–1314
    [Google Scholar]
  29. Redenbach M., Kieser H. M., Denapaite D., Eichner A., Cullum L., Kinashi H., Hopwood D. A. 1996; A set of ordered cosmids and a detailed genetic and physical map for the 8 Mb Streptomyces coelicolor A3(2) chromosome. Mol Microbiol 21:77–96 [CrossRef]
    [Google Scholar]
  30. 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]
  31. 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]
  32. Sekar V. 1987; A rapid screening procedure for identification of recombinant bacterial clones. Biotechniques 5:11–13
    [Google Scholar]
  33. Siddiquellah M., McGrath R., Vining L. C., Sala F., Westlake D. W. S. 1967; Biosynthesis of chloramphenicol II. p -Aminophenylalanine as a precursor of the p -nitrophenylserinol moiety. Can J Biochem 45:1881–1889 [CrossRef]
    [Google Scholar]
  34. Slock J., Stahly D. P., Han C. Y., Six E. W., Crawford I. P. 1990; An apparent Bacillus subtilis folic acid biosynthetic operon containing pab , an amphibolic trpG gene, a third gene required for synthesis of para-aminobenzoic acid, and the dihydropteroate synthase gene. J Bacteriol 172:7211–7226
    [Google Scholar]
  35. Southern E. M. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517 [CrossRef]
    [Google Scholar]
  36. Stuttard C. 1982; Temperate phages of Streptomyces venezuelae : lysogeny and host specificity shown by SV1 and SV2. J Gen Microbiol 128:115–121
    [Google Scholar]
  37. Swerdberg G., Skold O. 1980; Characterization of different plasmid-borne dihydropteroate synthases mediating bacterial resistance to sulfonamides. J Bacteriol 142:1–7
    [Google Scholar]
  38. Teng C. Y. P., Ganem B., Doktor S. Z., Nichol B. P., Bhatnagar R. K., Vining L. C. 1985; Total biosynthesis of 4-amino-4-deoxychorismic acid: a key intermediate in the biosynthesis of p -aminobenzoic acid and l- p -aminophenylalanine. J Am Chem Soc 107:5008–5009 [CrossRef]
    [Google Scholar]
  39. Thompson C. J., Skinner R. H., Thompson J., Ward J. M., Hopwood D. A., Cundliffe E. 1982; Biochemical characterization of resistance determinants cloned from antibiotic-producing streptomycetes. J Bacteriol 151:678–685
    [Google Scholar]
  40. Vats S., Stuttard C., Vining L. C. 1987; Transductional analysis of chloramphenicol biosynthesis genes in Streptomyces venezuelae. J Bacteriol 169:3809–3813
    [Google Scholar]
  41. Vining L. C., Stuttard C. 1994; Chloramphenicol. In Genetics and Biochemistry of Antibiotic Production pp 505–530 Edited by Vining L. C., Stuttard C. Boston: Butterworth-Heinemann;
    [Google Scholar]
  42. Wright F., Bibb M. J. 1992; Codon usage in the G+C-rich Streptomyces genome. Gene 113:55–65 [CrossRef]
    [Google Scholar]
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