Skip to main content
PMC home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1997 Jan;41(1):112–116. doi: 10.1128/aac.41.1.112

New tetracycline resistance determinants coding for ribosomal protection in streptococci and nucleotide sequence of tet(T) isolated from Streptococcus pyogenes A498.

D Clermont 1, O Chesneau 1, G De Cespédès 1, T Horaud 1
PMCID: PMC163670  PMID: 8980765

Abstract

An approach based on PCR has been developed to identify new members of the tet gene family in streptococci resistant to tetracycline and minocycline. Degenerate primers, corresponding to portions of the conserved domains of the proteins Tet(M), Tet(O), TeTB(P), Tet(Q), and Tet(S), all specifying the tetracycline-minocycline resistance phenotype, were used to selectively amplify DNA fragments within the coding sequences. Nine streptococcal strains which do not carry the genes tet(M), tet(O), tetB(P), tet(Q), or tet(S) were investigated. Four of them gave no detectable PCR products. The five remaining strains each yielded a PCR product of 1.1 kbp. DNA hybridization experiments showed that these putative Tet determinants fell into four new hybridization classes, of which one, Tet T, was further analyzed. The gene tet(T) was isolated from Streptococcus pyogenes A498, and the nucleotide sequence that was necessary and sufficient for the expression of tetracycline resistance in Escherichia coli was determined. The deduced Tet(T) protein consists of 651 amino acids. The protein most closely related to Tet(T) was Tet(Q), which has 49% identical amino acid residues. A phylogenetic analysis revealed that Tet T represents a novel branching order among the Tet determinants so far described.

Full Text

The Full Text of this article is available as a PDF (207.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allignet J., el Solh N. Diversity among the gram-positive acetyltransferases inactivating streptogramin A and structurally related compounds and characterization of a new staphylococcal determinant, vatB. Antimicrob Agents Chemother. 1995 Sep;39(9):2027–2036. doi: 10.1128/aac.39.9.2027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arthur M., Molinas C., Mabilat C., Courvalin P. Detection of erythromycin resistance by the polymerase chain reaction using primers in conserved regions of erm rRNA methylase genes. Antimicrob Agents Chemother. 1990 Oct;34(10):2024–2026. doi: 10.1128/aac.34.10.2024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bentorcha F., Clermont D., de Cespédès G., Horaud T. Natural occurrence of structures in oral streptococci and enterococci with DNA homology to Tn916. Antimicrob Agents Chemother. 1992 Jan;36(1):59–63. doi: 10.1128/aac.36.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burdett V. Nucleotide sequence of the tet(M) gene of Tn916. Nucleic Acids Res. 1990 Oct 25;18(20):6137–6137. doi: 10.1093/nar/18.20.6137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burdett V. Purification and characterization of Tet(M), a protein that renders ribosomes resistant to tetracycline. J Biol Chem. 1991 Feb 15;266(5):2872–2877. [PubMed] [Google Scholar]
  6. Charpentier E., Gerbaud G., Courvalin P. Characterization of a new class of tetracycline-resistance gene tet(S) in Listeria monocytogenes BM4210. Gene. 1993 Sep 6;131(1):27–34. doi: 10.1016/0378-1119(93)90665-p. [DOI] [PubMed] [Google Scholar]
  7. Chopra I., Hawkey P. M., Hinton M. Tetracyclines, molecular and clinical aspects. J Antimicrob Chemother. 1992 Mar;29(3):245–277. doi: 10.1093/jac/29.3.245. [DOI] [PubMed] [Google Scholar]
  8. Clermont D., Horaud T. Identification of chromosomal antibiotic resistance genes in Streptococcus anginosus ("S. milleri"). Antimicrob Agents Chemother. 1990 Sep;34(9):1685–1690. doi: 10.1128/aac.34.9.1685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. David F., de Céspèdes G., Delbos F., Horaud T. Diversity of chromosomal genetic elements and gene identification in antibiotic-resistant strains of Streptococcus pneumoniae and Streptococcus bovis. Plasmid. 1993 Mar;29(2):147–153. doi: 10.1006/plas.1993.1017. [DOI] [PubMed] [Google Scholar]
  10. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dittrich W., Schrempf H. The unstable tetracycline resistance gene of Streptomyces lividans 1326 encodes a putative protein with similarities to translational elongation factors and Tet(M) and Tet(O) proteins. Antimicrob Agents Chemother. 1992 May;36(5):1119–1124. doi: 10.1128/aac.36.5.1119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Doyle D., McDowall K. J., Butler M. J., Hunter I. S. Characterization of an oxytetracycline-resistance gene, otrA, of Streptomyces rimosus. Mol Microbiol. 1991 Dec;5(12):2923–2933. doi: 10.1111/j.1365-2958.1991.tb01852.x. [DOI] [PubMed] [Google Scholar]
  13. Gascoyne-Binzi D. M., Hawkey P. M., Heritage J. The distribution of variants of the Tet M determinant in tetracycline-resistant Neisseria gonorrhoeae. J Antimicrob Chemother. 1994 May;33(5):1011–1016. doi: 10.1093/jac/33.5.1011. [DOI] [PubMed] [Google Scholar]
  14. Guiney D. G., Hasegawa P., Bouic K., Matthews B. Genetic transfer systems in Bacteroides: cloning and mapping of the transferable tetracycline-resistance locus. Mol Microbiol. 1989 Nov;3(11):1617–1623. doi: 10.1111/j.1365-2958.1989.tb00147.x. [DOI] [PubMed] [Google Scholar]
  15. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  16. Higgins D. G., Bleasby A. J., Fuchs R. CLUSTAL V: improved software for multiple sequence alignment. Comput Appl Biosci. 1992 Apr;8(2):189–191. doi: 10.1093/bioinformatics/8.2.189. [DOI] [PubMed] [Google Scholar]
  17. Le Bouguénec C., de Cespédès G., Horaud T. Molecular analysis of a composite chromosomal conjugative element (Tn3701) of Streptococcus pyogenes. J Bacteriol. 1988 Sep;170(9):3930–3936. doi: 10.1128/jb.170.9.3930-3936.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. LeBlanc D. J., Lee L. N., Titmas B. M., Smith C. J., Tenover F. C. Nucleotide sequence analysis of tetracycline resistance gene tetO from Streptococcus mutans DL5. J Bacteriol. 1988 Aug;170(8):3618–3626. doi: 10.1128/jb.170.8.3618-3626.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lépine G., Lacroix J. M., Walker C. B., Progulske-Fox A. Sequencing of a tet(Q) gene isolated from Bacteroides fragilis 1126. Antimicrob Agents Chemother. 1993 Sep;37(9):2037–2041. doi: 10.1128/aac.37.9.2037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Manavathu E. K., Fernandez C. L., Cooperman B. S., Taylor D. E. Molecular studies on the mechanism of tetracycline resistance mediated by Tet(O). Antimicrob Agents Chemother. 1990 Jan;34(1):71–77. doi: 10.1128/aac.34.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Manavathu E. K., Hiratsuka K., Taylor D. E. Nucleotide sequence analysis and expression of a tetracycline-resistance gene from Campylobacter jejuni. Gene. 1988;62(1):17–26. doi: 10.1016/0378-1119(88)90576-8. [DOI] [PubMed] [Google Scholar]
  22. Martin P., Trieu-Cuot P., Courvalin P. Nucleotide sequence of the tetM tetracycline resistance determinant of the streptococcal conjugative shuttle transposon Tn1545. Nucleic Acids Res. 1986 Sep 11;14(17):7047–7058. doi: 10.1093/nar/14.17.7047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McLaughlin J. R., Murray C. L., Rabinowitz J. C. Unique features in the ribosome binding site sequence of the gram-positive Staphylococcus aureus beta-lactamase gene. J Biol Chem. 1981 Nov 10;256(21):11283–11291. [PubMed] [Google Scholar]
  24. Moran C. P., Jr, Lang N., LeGrice S. F., Lee G., Stephens M., Sonenshein A. L., Pero J., Losick R. Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet. 1982;186(3):339–346. doi: 10.1007/BF00729452. [DOI] [PubMed] [Google Scholar]
  25. Nesin M., Svec P., Lupski J. R., Godson G. N., Kreiswirth B., Kornblum J., Projan S. J. Cloning and nucleotide sequence of a chromosomally encoded tetracycline resistance determinant, tetA(M), from a pathogenic, methicillin-resistant strain of Staphylococcus aureus. Antimicrob Agents Chemother. 1990 Nov;34(11):2273–2276. doi: 10.1128/aac.34.11.2273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nikolich M. P., Hong G., Shoemaker N. B., Salyers A. A. Evidence for natural horizontal transfer of tetQ between bacteria that normally colonize humans and bacteria that normally colonize livestock. Appl Environ Microbiol. 1994 Sep;60(9):3255–3260. doi: 10.1128/aem.60.9.3255-3260.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nikolich M. P., Shoemaker N. B., Salyers A. A. A Bacteroides tetracycline resistance gene represents a new class of ribosome protection tetracycline resistance. Antimicrob Agents Chemother. 1992 May;36(5):1005–1012. doi: 10.1128/aac.36.5.1005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pang Y., Brown B. A., Steingrube V. A., Wallace R. J., Jr, Roberts M. C. Tetracycline resistance determinants in Mycobacterium and Streptomyces species. Antimicrob Agents Chemother. 1994 Jun;38(6):1408–1412. doi: 10.1128/aac.38.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ridenhour M. B., Fletcher H. M., Mortensen J. E., Daneo-Moore L. A novel tetracycline-resistant determinant, tet(U), is encoded on the plasmid pKq10 in Enterococcus faecium. Plasmid. 1996 Mar;35(2):71–80. doi: 10.1006/plas.1996.0009. [DOI] [PubMed] [Google Scholar]
  31. Roberts M. C. Epidemiology of tetracycline-resistance determinants. Trends Microbiol. 1994 Oct;2(10):353–357. doi: 10.1016/0966-842x(94)90610-6. [DOI] [PubMed] [Google Scholar]
  32. Saitou N., Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987 Jul;4(4):406–425. doi: 10.1093/oxfordjournals.molbev.a040454. [DOI] [PubMed] [Google Scholar]
  33. Sanchez-Pescador R., Brown J. T., Roberts M., Urdea M. S. Homology of the TetM with translational elongation factors: implications for potential modes of tetM-conferred tetracycline resistance. Nucleic Acids Res. 1988 Feb 11;16(3):1218–1218. doi: 10.1093/nar/16.3.1218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sanchez-Pescador R., Brown J. T., Roberts M., Urdea M. S. The nucleotide sequence of the tetracycline resistance determinant tetM from Ureaplasma urealyticum. Nucleic Acids Res. 1988 Feb 11;16(3):1216–1217. doi: 10.1093/nar/16.3.1216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Sloan J., McMurry L. M., Lyras D., Levy S. B., Rood J. I. The Clostridium perfringens Tet P determinant comprises two overlapping genes: tetA(P), which mediates active tetracycline efflux, and tetB(P), which is related to the ribosomal protection family of tetracycline-resistance determinants. Mol Microbiol. 1994 Jan;11(2):403–415. doi: 10.1111/j.1365-2958.1994.tb00320.x. [DOI] [PubMed] [Google Scholar]
  37. Su Y. A., He P., Clewell D. B. Characterization of the tet(M) determinant of Tn916: evidence for regulation by transcription attenuation. Antimicrob Agents Chemother. 1992 Apr;36(4):769–778. doi: 10.1128/aac.36.4.769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Taylor D. E., Chau A. Tetracycline resistance mediated by ribosomal protection. Antimicrob Agents Chemother. 1996 Jan;40(1):1–5. doi: 10.1128/aac.40.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Taylor D. E., Hiratsuka K., Ray H., Manavathu E. K. Characterization and expression of a cloned tetracycline resistance determinant from Campylobacter jejuni plasmid pUA466. J Bacteriol. 1987 Jul;169(7):2984–2989. doi: 10.1128/jb.169.7.2984-2989.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
  41. Trieu-Cuot P., de Cespédès G., Bentorcha F., Delbos F., Gaspar E., Horaud T. Study of heterogeneity of chloramphenicol acetyltransferase (CAT) genes in streptococci and enterococci by polymerase chain reaction: characterization of a new CAT determinant. Antimicrob Agents Chemother. 1993 Dec;37(12):2593–2598. doi: 10.1128/aac.37.12.2593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wang Y., Taylor D. E. A DNA sequence upstream of the tet(O) gene is required for full expression of tetracycline resistance. Antimicrob Agents Chemother. 1991 Oct;35(10):2020–2025. doi: 10.1128/aac.35.10.2020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Yakhnin A. V., Vorozheykina D. P., Matvienko N. I. Nucleotide sequence of the Thermus thermophilus HB8 gene coding for elongation factor G. Nucleic Acids Res. 1989 Nov 11;17(21):8863–8863. doi: 10.1093/nar/17.21.8863. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES