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. 1992 Oct;58(10):3399–3403. doi: 10.1128/aem.58.10.3399-3403.1992

Cloning and expression of the aspartate carbamoyltransferase gene from Treponema denticola.

K Ishihara 1, M Ishihara 1, I Takazoe 1, K Okuda 1
PMCID: PMC183110  PMID: 1444372

Abstract

Treponema denticola seems to play a central role in the etiology of human periodontal disease. We have cloned an antigenic protein-coding sequence from T. denticola ATCC 33520. The protein-coding region was found to be a 3-kbp HindIII-HindIII fragment. The open reading frame consists of 1,426 bp and codes for a protein with an M(r) of 54,919. The deduced amino acid sequence showed 33.8% homology with that of the aspartate carbamoyltransferase of Escherichia coli. The gene products showed aspartate carbamoyltransferase activity.

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Selected References

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  1. Armitage G. C., Dickinson W. R., Jenderseck R. S., Levine S. M., Chambers D. W. Relationship between the percentage of subgingival spirochetes and the severity of periodontal disease. J Periodontol. 1982 Sep;53(9):550–556. doi: 10.1902/jop.1982.53.9.550. [DOI] [PubMed] [Google Scholar]
  2. Bencini D. A., Houghton J. E., Hoover T. A., Foltermann K. F., Wild J. R., O'Donovan G. A. The DNA sequence of argI from Escherichia coli K12. Nucleic Acids Res. 1983 Dec 10;11(23):8509–8518. doi: 10.1093/nar/11.23.8509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boehringer H., Berthold P. H., Taichman N. S. Studies on the interaction of human neutrophils with plaque spirochetes. J Periodontal Res. 1986 May;21(3):195–209. doi: 10.1111/j.1600-0765.1986.tb01452.x. [DOI] [PubMed] [Google Scholar]
  4. Boehringer H., Taichman N. S., Shenker B. J. Suppression of fibroblast proliferation by oral spirochetes. Infect Immun. 1984 Jul;45(1):155–159. doi: 10.1128/iai.45.1.155-159.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  6. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gold L., Pribnow D., Schneider T., Shinedling S., Singer B. S., Stormo G. Translational initiation in prokaryotes. Annu Rev Microbiol. 1981;35:365–403. doi: 10.1146/annurev.mi.35.100181.002053. [DOI] [PubMed] [Google Scholar]
  8. Gouaux J. E., Lipscomb W. N. Three-dimensional structure of carbamoyl phosphate and succinate bound to aspartate carbamoyltransferase. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4205–4208. doi: 10.1073/pnas.85.12.4205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Grenier D. Characteristics of hemolytic and hemagglutinating activities of Treponema denticola. Oral Microbiol Immunol. 1991 Aug;6(4):246–249. doi: 10.1111/j.1399-302x.1991.tb00485.x. [DOI] [PubMed] [Google Scholar]
  10. Grenier D., Uitto V. J., McBride B. C. Cellular location of a Treponema denticola chymotrypsinlike protease and importance of the protease in migration through the basement membrane. Infect Immun. 1990 Feb;58(2):347–351. doi: 10.1128/iai.58.2.347-351.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hoover T. A., Roof W. D., Foltermann K. F., O'Donovan G. A., Bencini D. A., Wild J. R. Nucleotide sequence of the structural gene (pyrB) that encodes the catalytic polypeptide of aspartate transcarbamoylase of Escherichia coli. Proc Natl Acad Sci U S A. 1983 May;80(9):2462–2466. doi: 10.1073/pnas.80.9.2462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. JOHNSON R. C., ROGERS P. 5-FLUOROURACIL AS A SELECTIVE AGENT FOR GROWTH OF LEPTOSPIRAE. J Bacteriol. 1964 Feb;87:422–426. doi: 10.1128/jb.87.2.422-426.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. JOHNSON R. C., ROGERS P. METABOLISM OF LEPTOSPIRAE. I. UTILIZATION OF AMINO ACIDS AND PURINE, AND PYRIMIDINE BASES. Arch Biochem Biophys. 1964 Sep;107:459–470. doi: 10.1016/0003-9861(64)90302-9. [DOI] [PubMed] [Google Scholar]
  14. Krause K. L., Volz K. W., Lipscomb W. N. 2.5 A structure of aspartate carbamoyltransferase complexed with the bisubstrate analog N-(phosphonacetyl)-L-aspartate. J Mol Biol. 1987 Feb 5;193(3):527–553. doi: 10.1016/0022-2836(87)90265-8. [DOI] [PubMed] [Google Scholar]
  15. LISTGARTEN M. A. ELECTRON MICROSCOPIC OBSERVATIONS ON THE BACTERIAL FLORA OF ACUTE NECROTIZING ULCERATIVE GINGIVITIS. J Periodontol. 1965 Jul-Aug;36:328–339. doi: 10.1902/jop.1965.36.4.328. [DOI] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Lerner C. G., Switzer R. L. Cloning and structure of the Bacillus subtilis aspartate transcarbamylase gene (pyrB). J Biol Chem. 1986 Aug 25;261(24):11156–11165. [PubMed] [Google Scholar]
  18. Loenen W. A., Brammar W. J. A bacteriophage lambda vector for cloning large DNA fragments made with several restriction enzymes. Gene. 1980 Aug;10(3):249–259. doi: 10.1016/0378-1119(80)90054-2. [DOI] [PubMed] [Google Scholar]
  19. Loesche W. J., Syed S. A., Schmidt E., Morrison E. C. Bacterial profiles of subgingival plaques in periodontitis. J Periodontol. 1985 Aug;56(8):447–456. doi: 10.1902/jop.1985.56.8.447. [DOI] [PubMed] [Google Scholar]
  20. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  21. Michaels G., Kelln R. A., Nargang F. E. Cloning, nucleotide sequence and expression of the pyrBI operon of Salmonella typhimurium LT2. Eur J Biochem. 1987 Jul 1;166(1):55–61. doi: 10.1111/j.1432-1033.1987.tb13483.x. [DOI] [PubMed] [Google Scholar]
  22. Miyamoto M., Noji S., Kokeguchi S., Kato K., Kurihara H., Murayama Y., Taniguchi S. Molecular cloning and sequence analysis of antigen gene tdpA of Treponema denticola. Infect Immun. 1991 Jun;59(6):1941–1947. doi: 10.1128/iai.59.6.1941-1947.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ohta K., Makinen K. K., Loesche W. J. Purification and characterization of an enzyme produced by Treponema denticola capable of hydrolyzing synthetic trypsin substrates. Infect Immun. 1986 Jul;53(1):213–220. doi: 10.1128/iai.53.1.213-220.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Potvin B. W., Kelleher R. J., Jr, Gooder H. Pyrimidine biosynthetic pathway of Baccillus subtilis. J Bacteriol. 1975 Aug;123(2):604–615. doi: 10.1128/jb.123.2.604-615.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Prescott L. M., Jones M. E. Modified methods for the determination of carbamyl aspartate. Anal Biochem. 1969 Dec;32(3):408–419. doi: 10.1016/s0003-2697(69)80008-4. [DOI] [PubMed] [Google Scholar]
  26. Que X. C., Kuramitsu H. K. Isolation and characterization of the Treponema denticola prtA gene coding for chymotrypsinlike protease activity and detection of a closely linked gene encoding PZ-PLGPA-hydrolyzing activity. Infect Immun. 1990 Dec;58(12):4099–4105. doi: 10.1128/iai.58.12.4099-4105.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schachman H. K., Pauza C. D., Navre M., Karels M. J., Wu L., Yang Y. R. Location of amino acid alterations in mutants of aspartate transcarbamoylase: Structural aspects of interallelic complementation. Proc Natl Acad Sci U S A. 1984 Jan;81(1):115–119. doi: 10.1073/pnas.81.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sela M. N., Weinberg A., Borinsky R., Holt S. C., Dishon T. Inhibition of superoxide production in human polymorphonuclear leukocytes by oral treponemal factors. Infect Immun. 1988 Mar;56(3):589–594. doi: 10.1128/iai.56.3.589-594.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Shenker B. J., Listgarten M. A., Taichman N. S. Suppression of human lymphocyte responses by oral spirochetes: a monocyte-dependent phenomenon. J Immunol. 1984 Apr;132(4):2039–2045. [PubMed] [Google Scholar]
  31. Shigesada K., Stark G. R., Maley J. A., Niswander L. A., Davidson J. N. Construction of a cDNA to the hamster CAD gene and its application toward defining the domain for aspartate transcarbamylase. Mol Cell Biol. 1985 Jul;5(7):1735–1742. doi: 10.1128/mcb.5.7.1735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Simonson L. G., Goodman C. H., Morton H. E. Quantitative immunoassay of Treponema denticola serovar C in adult periodontitis. J Clin Microbiol. 1990 Jul;28(7):1493–1496. doi: 10.1128/jcm.28.7.1493-1496.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Van Vliet F., Cunin R., Jacobs A., Piette J., Gigot D., Lauwereys M., Piérard A., Glansdorff N. Evolutionary divergence of genes for ornithine and aspartate carbamoyl-transferases--complete sequence and mode of regulation of the Escherichia coli argF gene; comparison of argF with argI and pyrB. Nucleic Acids Res. 1984 Aug 10;12(15):6277–6289. doi: 10.1093/nar/12.15.6277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]

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