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. 1992 Oct;174(20):6563–6570. doi: 10.1128/jb.174.20.6563-6570.1992

Tryptophan biosynthesis genes in Lactococcus lactis subsp. lactis.

J Bardowski 1, S D Ehrlich 1, A Chopin 1
PMCID: PMC207625  PMID: 1400208

Abstract

The Lactococcus lactis chromosomal region containing the seven structural genes required for tryptophan biosynthesis was characterized by cloning and sequencing. All of the trp genes were identified by the homology of their products with known Trp proteins from other organisms. The identification was confirmed for five genes by their ability to complement trp mutations in Escherichia coli. The seven structural genes are present in the order trpEGDCFBA and span a 7,968-bp segment. Each gene is preceded by a putative ribosome binding site complementary to the 3' end of the L. lactis 16S rRNA. Three pairs of genes (trpG-trpD, trpC-trpF, and trpB-trpA) overlap, and there is intercistronic spacing of 124, 46, and 585 bp between the trpE-trpG, trpD-trpC, and trpF-trpB gene pairs, respectively. No gene fusion was found. Upstream of the trp genes, a 457-bp noncoding DNA segment contains several regions fitting the consensus for gram-positive promoters and one region strongly resembling a transcription terminator. However, it seems unlikely that an attenuation mechanism similar to the one found in E. coli regulates tryptophan biosynthesis in L. lactis, since no potential leader peptide was detected. We propose that a mechanisms resembling that described in Bacillus spp. can regulate trp genes expression in L. lactis.

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Selected References

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  1. Anagnostopoulos C., Spizizen J. REQUIREMENTS FOR TRANSFORMATION IN BACILLUS SUBTILIS. J Bacteriol. 1961 May;81(5):741–746. doi: 10.1128/jb.81.5.741-746.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bae Y. M., Holmgren E., Crawford I. P. Rhizobium meliloti anthranilate synthase gene: cloning, sequence, and expression in Escherichia coli. J Bacteriol. 1989 Jun;171(6):3471–3478. doi: 10.1128/jb.171.6.3471-3478.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Band L., Shimotsu H., Henner D. J. Nucleotide sequence of the Bacillus subtilis trpE and trpD genes. Gene. 1984 Jan;27(1):55–65. doi: 10.1016/0378-1119(84)90238-5. [DOI] [PubMed] [Google Scholar]
  4. Calhoun D. H., Pierson D. L., Jensen R. A. The regulation of tryptophan biosynthesis in Pseudomonas aeruginosa. Mol Gen Genet. 1973 Mar 1;121(2):117–132. doi: 10.1007/BF00277526. [DOI] [PubMed] [Google Scholar]
  5. Chang M., Hadero A., Crawford I. P. Sequence of the Pseudomonas aeruginosa trpI activator gene and relatedness of trpI to other procaryotic regulatory genes. J Bacteriol. 1989 Jan;171(1):172–183. doi: 10.1128/jb.171.1.172-183.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chopin A., Chopin M. C., Moillo-Batt A., Langella P. Two plasmid-determined restriction and modification systems in Streptococcus lactis. Plasmid. 1984 May;11(3):260–263. doi: 10.1016/0147-619x(84)90033-7. [DOI] [PubMed] [Google Scholar]
  7. Cluzel P. J., Chopin A., Ehrlich S. D., Chopin M. C. Phage abortive infection mechanism from Lactococcus lactis subsp. lactis, expression of which is mediated by an Iso-ISS1 element. Appl Environ Microbiol. 1991 Dec;57(12):3547–3551. doi: 10.1128/aem.57.12.3547-3551.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Corpet F. Multiple sequence alignment with hierarchical clustering. Nucleic Acids Res. 1988 Nov 25;16(22):10881–10890. doi: 10.1093/nar/16.22.10881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Crawford I. P. Evolution of a biosynthetic pathway: the tryptophan paradigm. Annu Rev Microbiol. 1989;43:567–600. doi: 10.1146/annurev.mi.43.100189.003031. [DOI] [PubMed] [Google Scholar]
  10. Crawford I. P. Gene rearrangements in the evolution of the tryptophan synthetic pathway. Bacteriol Rev. 1975 Jun;39(2):87–120. doi: 10.1128/br.39.2.87-120.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Crawford I. P., Gunsalus I. C. Inducibility of tryptophan synthetase in Pseudomonas putida. Proc Natl Acad Sci U S A. 1966 Aug;56(2):717–724. doi: 10.1073/pnas.56.2.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Delorme C., Ehrlich S. D., Renault P. Histidine biosynthesis genes in Lactococcus lactis subsp. lactis. J Bacteriol. 1992 Oct;174(20):6571–6579. doi: 10.1128/jb.174.20.6571-6579.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dower W. J., Miller J. F., Ragsdale C. W. High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res. 1988 Jul 11;16(13):6127–6145. doi: 10.1093/nar/16.13.6127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Essar D. W., Eberly L., Han C. Y., Crawford I. P. DNA sequences and characterization of four early genes of the tryptophan pathway in Pseudomonas aeruginosa. J Bacteriol. 1990 Feb;172(2):853–866. doi: 10.1128/jb.172.2.853-866.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fickett J. W. Recognition of protein coding regions in DNA sequences. Nucleic Acids Res. 1982 Sep 11;10(17):5303–5318. doi: 10.1093/nar/10.17.5303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Furter R., Paravicini G., Aebi M., Braus G., Prantl F., Niederberger P., Hütter R. The TRP4 gene of Saccharomyces cerevisiae: isolation and structural analysis. Nucleic Acids Res. 1986 Aug 26;14(16):6357–6373. doi: 10.1093/nar/14.16.6357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Godon J. J., Chopin M. C., Ehrlich S. D. Branched-chain amino acid biosynthesis genes in Lactococcus lactis subsp. lactis. J Bacteriol. 1992 Oct;174(20):6580–6589. doi: 10.1128/jb.174.20.6580-6589.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gollnick P., Ishino S., Kuroda M. I., Henner D. J., Yanofsky C. The mtr locus is a two-gene operon required for transcription attenuation in the trp operon of Bacillus subtilis. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8726–8730. doi: 10.1073/pnas.87.22.8726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hadero A., Crawford I. P. Nucleotide sequence of the genes for tryptophan synthase in Pseudomonas aeruginosa. Mol Biol Evol. 1986 May;3(3):191–204. doi: 10.1093/oxfordjournals.molbev.a040388. [DOI] [PubMed] [Google Scholar]
  20. Henner D. J., Band L., Shimotsu H. Nucleotide sequence of the Bacillus subtilis tryptophan operon. Gene. 1985;34(2-3):169–177. doi: 10.1016/0378-1119(85)90125-8. [DOI] [PubMed] [Google Scholar]
  21. Hill C., Miller L. A., Klaenhammer T. R. In vivo genetic exchange of a functional domain from a type II A methylase between lactococcal plasmid pTR2030 and a virulent bacteriophage. J Bacteriol. 1991 Jul;173(14):4363–4370. doi: 10.1128/jb.173.14.4363-4370.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hill C., Miller L. A., Klaenhammer T. R. Nucleotide sequence and distribution of the pTR2030 resistance determinant (hsp) which aborts bacteriophage infection in lactococci. Appl Environ Microbiol. 1990 Jul;56(7):2255–2258. doi: 10.1128/aem.56.7.2255-2258.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kaletta C., Entian K. D. Nisin, a peptide antibiotic: cloning and sequencing of the nisA gene and posttranslational processing of its peptide product. J Bacteriol. 1989 Mar;171(3):1597–1601. doi: 10.1128/jb.171.3.1597-1601.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Koivula T., Hemilä H. Nucleotide sequence of a Lactococcus lactis gene cluster encoding adenylate kinase, initiation factor 1 and ribosomal proteins. J Gen Microbiol. 1991 Nov;137(11):2595–2600. doi: 10.1099/00221287-137-11-2595. [DOI] [PubMed] [Google Scholar]
  25. Koivula T., Palva I., Hemilä H. Nucleotide sequence of the secY gene from Lactococcus lactis and identification of conserved regions by comparison of four SecY proteins. FEBS Lett. 1991 Aug 19;288(1-2):114–118. doi: 10.1016/0014-5793(91)81015-z. [DOI] [PubMed] [Google Scholar]
  26. Kok J., Leenhouts K. J., Haandrikman A. J., Ledeboer A. M., Venema G. Nucleotide sequence of the cell wall proteinase gene of Streptococcus cremoris Wg2. Appl Environ Microbiol. 1988 Jan;54(1):231–238. doi: 10.1128/aem.54.1.231-238.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kuroda M. I., Henner D., Yanofsky C. cis-acting sites in the transcript of the Bacillus subtilis trp operon regulate expression of the operon. J Bacteriol. 1988 Jul;170(7):3080–3088. doi: 10.1128/jb.170.7.3080-3088.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kuroda M. I., Shimotsu H., Henner D. J., Yanofsky C. Regulatory elements common to the Bacillus pumilus and Bacillus subtilis trp operons. J Bacteriol. 1986 Sep;167(3):792–798. doi: 10.1128/jb.167.3.792-798.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lam W. L., Cohen A., Tsouluhas D., Doolittle W. F. Genes for tryptophan biosynthesis in the archaebacterium Haloferax volcanii. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6614–6618. doi: 10.1073/pnas.87.17.6614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Le Bourgeois P., Mata M., Ritzenthaler P. Genome comparison of Lactococcus strains by pulsed-field gel electrophoresis. FEMS Microbiol Lett. 1989 May;50(1-2):65–69. doi: 10.1016/0378-1097(89)90460-6. [DOI] [PubMed] [Google Scholar]
  31. Matsui K., Sano K., Ohtsubo E. Complete nucleotide and deduced amino acid sequences of the Brevibacterium lactofermentum tryptophan operon. Nucleic Acids Res. 1986 Dec 22;14(24):10113–10114. doi: 10.1093/nar/14.24.10113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Mayo B., Kok J., Venema K., Bockelmann W., Teuber M., Reinke H., Venema G. Molecular cloning and sequence analysis of the X-prolyl dipeptidyl aminopeptidase gene from Lactococcus lactis subsp. cremoris. Appl Environ Microbiol. 1991 Jan;57(1):38–44. doi: 10.1128/aem.57.1.38-44.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Meile L., Stettler R., Banholzer R., Kotik M., Leisinger T. Tryptophan gene cluster of Methanobacterium thermoautotrophicum Marburg: molecular cloning and nucleotide sequence of a putative trpEGCFBAD operon. J Bacteriol. 1991 Aug;173(16):5017–5023. doi: 10.1128/jb.173.16.5017-5023.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Nardi M., Chopin M. C., Chopin A., Cals M. M., Gripon J. C. Cloning and DNA sequence analysis of an X-prolyl dipeptidyl aminopeptidase gene from Lactococcus lactis subsp. lactis NCDO 763. Appl Environ Microbiol. 1991 Jan;57(1):45–50. doi: 10.1128/aem.57.1.45-50.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Natori Y., Kano Y., Imamoto F. Nucleotide sequences and genomic constitution of five tryptophan genes of Lactobacillus casei. J Biochem. 1990 Feb;107(2):248–255. doi: 10.1093/oxfordjournals.jbchem.a123034. [DOI] [PubMed] [Google Scholar]
  36. Renault P., Gaillardin C., Heslot H. Product of the Lactococcus lactis gene required for malolactic fermentation is homologous to a family of positive regulators. J Bacteriol. 1989 Jun;171(6):3108–3114. doi: 10.1128/jb.171.6.3108-3114.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sano K., Matsui K. Structure and function of the trp operon control regions of Brevibacterium lactofermentum, a glutamic-acid-producing bacterium. Gene. 1987;53(2-3):191–200. doi: 10.1016/0378-1119(87)90007-2. [DOI] [PubMed] [Google Scholar]
  38. Sato S., Nakada Y., Kanaya S., Tanaka T. Molecular cloning and nucleotide sequence of Thermus thermophilus HB8 trpE and trpG. Biochim Biophys Acta. 1988 Sep 7;950(3):303–312. doi: 10.1016/0167-4781(88)90126-1. [DOI] [PubMed] [Google Scholar]
  39. Shimotsu H., Kuroda M. I., Yanofsky C., Henner D. J. Novel form of transcription attenuation regulates expression the Bacillus subtilis tryptophan operon. J Bacteriol. 1986 May;166(2):461–471. doi: 10.1128/jb.166.2.461-471.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sibold L., Henriquet M. Cloning of the trp genes from the archaebacterium Methanococcus voltae: nucleotide sequence of the trpBA genes. Mol Gen Genet. 1988 Nov;214(3):439–450. doi: 10.1007/BF00330478. [DOI] [PubMed] [Google Scholar]
  41. Simon D., Chopin A. Construction of a vector plasmid family and its use for molecular cloning in Streptococcus lactis. Biochimie. 1988 Apr;70(4):559–566. doi: 10.1016/0300-9084(88)90093-4. [DOI] [PubMed] [Google Scholar]
  42. Slock J., Stahly D. P., Han C. Y., Six E. W., Crawford I. P. 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. 1990 Dec;172(12):7211–7226. doi: 10.1128/jb.172.12.7211-7226.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tanaka T. Restriction of plasmid-mediated transformation in Bacillus subtilis 168. Mol Gen Genet. 1979 Sep;175(2):235–237. doi: 10.1007/BF00425542. [DOI] [PubMed] [Google Scholar]
  44. Terzaghi B. E., Sandine W. E. Improved medium for lactic streptococci and their bacteriophages. Appl Microbiol. 1975 Jun;29(6):807–813. doi: 10.1128/am.29.6.807-813.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tschumper G., Carbon J. Sequence of a yeast DNA fragment containing a chromosomal replicator and the TRP1 gene. Gene. 1980 Jul;10(2):157–166. doi: 10.1016/0378-1119(80)90133-x. [DOI] [PubMed] [Google Scholar]
  46. Yanofsky C. Comparison of regulatory and structural regions of genes of tryptophan metabolism. Mol Biol Evol. 1984 Feb;1(2):143–161. doi: 10.1093/oxfordjournals.molbev.a040307. [DOI] [PubMed] [Google Scholar]
  47. Yanofsky C., Platt T., Crawford I. P., Nichols B. P., Christie G. E., Horowitz H., VanCleemput M., Wu A. M. The complete nucleotide sequence of the tryptophan operon of Escherichia coli. Nucleic Acids Res. 1981 Dec 21;9(24):6647–6668. doi: 10.1093/nar/9.24.6647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Zalkin H., Paluh J. L., van Cleemput M., Moye W. S., Yanofsky C. Nucleotide sequence of Saccharomyces cerevisiae genes TRP2 and TRP3 encoding bifunctional anthranilate synthase: indole-3-glycerol phosphate synthase. J Biol Chem. 1984 Mar 25;259(6):3985–3992. [PubMed] [Google Scholar]
  49. Zalkin H., Yanofsky C. Yeast gene TRP5: structure, function, regulation. J Biol Chem. 1982 Feb 10;257(3):1491–1500. [PubMed] [Google Scholar]
  50. d'Aubenton Carafa Y., Brody E., Thermes C. Prediction of rho-independent Escherichia coli transcription terminators. A statistical analysis of their RNA stem-loop structures. J Mol Biol. 1990 Dec 20;216(4):835–858. doi: 10.1016/s0022-2836(99)80005-9. [DOI] [PubMed] [Google Scholar]
  51. de Vos W. M., Boerrigter I., van Rooyen R. J., Reiche B., Hengstenberg W. Characterization of the lactose-specific enzymes of the phosphotransferase system in Lactococcus lactis. J Biol Chem. 1990 Dec 25;265(36):22554–22560. [PubMed] [Google Scholar]
  52. te Riele H., Michel B., Ehrlich S. D. Single-stranded plasmid DNA in Bacillus subtilis and Staphylococcus aureus. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2541–2545. doi: 10.1073/pnas.83.8.2541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. van Asseldonk M., Rutten G., Oteman M., Siezen R. J., de Vos W. M., Simons G. Cloning of usp45, a gene encoding a secreted protein from Lactococcus lactis subsp. lactis MG1363. Gene. 1990 Oct 30;95(1):155–160. doi: 10.1016/0378-1119(90)90428-t. [DOI] [PubMed] [Google Scholar]
  54. van Belkum M. J., Hayema B. J., Jeeninga R. E., Kok J., Venema G. Organization and nucleotide sequences of two lactococcal bacteriocin operons. Appl Environ Microbiol. 1991 Feb;57(2):492–498. doi: 10.1128/aem.57.2.492-498.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. van Rooijen R. J., de Vos W. M. Molecular cloning, transcriptional analysis, and nucleotide sequence of lacR, a gene encoding the repressor of the lactose phosphotransferase system of Lactococcus lactis. J Biol Chem. 1990 Oct 25;265(30):18499–18503. [PubMed] [Google Scholar]
  56. van Rooijen R. J., van Schalkwijk S., de Vos W. M. Molecular cloning, characterization, and nucleotide sequence of the tagatose 6-phosphate pathway gene cluster of the lactose operon of Lactococcus lactis. J Biol Chem. 1991 Apr 15;266(11):7176–7181. [PubMed] [Google Scholar]

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