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PMID: 11717269 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Transcriptional analysis of the tet(P) operon from Clostridium perfringens.

Journal of bacteriology ·Vol. 183 ·No. 24 ·2001-12-00 ·Pages 7110-9

Johanesen PA, Lyras D, Bannam TL, Rood JI

Abstract

The Clostridium perfringens tetracycline resistance determinant from the 47-kb conjugative R-plasmid pCW3 is unique in that it consists of two overlapping genes, tetA(P) and tetB(P), which mediate resistance by different mechanisms. Detailed transcriptional analysis has shown that the inducible tetA(P) and tetB(P) genes comprise an operon that is transcribed from a single promoter, P3, located 529 bp upstream of the tetA(P) start codon. Deletion of P3 or alteration of the spacing between the -35 and -10 regions significantly reduced the level of transcription in a reporter construct. Induction was shown to be mediated at the level of transcription. Unexpectedly, a factor-independent terminator, T1, was detected downstream of P3 but before the start of the tetA(P) gene. Deletion or mutation of this terminator led to increased read-through transcription in the reporter construct. It is postulated that the T1 terminator is an intrinsic control element of the tet(P) operon and that it acts to prevent the overexpression of the TetA(P) transmembrane protein, even in the presence of tetracycline.

MeSH Terms
Antiporters/genetics Bacterial Proteins/genetics Base Sequence Clostridium perfringens/genetics Gene Expression Regulation, Bacterial Genes, Reporter Molecular Sequence Data Nucleic Acid Conformation Operon/genetics Promoter Regions, Genetic RNA, Bacterial Recombinant Fusion Proteins Terminator Regions, Genetic Tetracycline Resistance/genetics Transcription, Genetic
Chemicals
Antiporters Bacterial Proteins RNA, Bacterial Recombinant Fusion Proteins TETP protein, Clostridium perfringens tetA protein, Bacteria
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Johanesen P A
Bacterial Pathogenesis Research Group, Department of Microbiology, Monash University, Victoria 3800, Australia.
Lyras D
Bannam T L
Rood J I
References (47)
47 references, click to expand
  1. Nucleotide sequence of the tetracycline resistance gene of pTHT15, a thermophilic Bacillus plasmid: comparison with staphylococcal TcR controls.
    Gene. 1985;37(1-3):131-8 PMID: 2996983
  2. Worldwide distribution of the conjugative Clostridium perfringens tetracycline resistance plasmid, pCW3.
    Plasmid. 1985 Jul;14(1):37-46 PMID: 2863833
  3. Analysis of E. coli promoter sequences.
    Nucleic Acids Res. 1987 Mar 11;15(5):2343-61 PMID: 3550697
  4. Hybridization analysis of the class P tetracycline resistance determinant from the Clostridium perfringens R-plasmid, pCW3.
    Plasmid. 1988 Mar;19(2):113-20 PMID: 2901767
  5. Characterization of the tetracycline resistance gene of plasmid pT181 of Staphylococcus aureus.
    J Bacteriol. 1988 Dec;170(12):5522-8 PMID: 3142848
  6. Nucleotide sequence analysis and expression studies of a chloramphenicol-acetyltransferase-coding gene from Clostridium perfringens.
    Gene. 1989 Feb 20;75(2):349-54 PMID: 2541053
  7. Overproduction of transposon Tn10-encoded tetracycline resistance protein results in cell death and loss of membrane potential.
    J Bacteriol. 1989 Jun;171(6):3557-9 PMID: 2542231
  8. Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension.
    Gene. 1989 Apr 15;77(1):61-8 PMID: 2744488
  9. Promoter recognition by Escherichia coli RNA polymerase. Role of the spacer DNA in functional complex formation.
    J Mol Biol. 1989 Jun 20;207(4):749-56 PMID: 2668539
  10. Electroporation-mediated transformation of lysostaphin-treated Clostridium perfringens.
    Gene. 1989 Oct 30;82(2):327-33 PMID: 2555265
  11. Computer prediction of RNA structure.
    Methods Enzymol. 1989;180:262-88 PMID: 2482418
  12. Effects of carriage and expression of the Tn10 tetracycline-resistance operon on the fitness of Escherichia coli K12.
    Mol Biol Evol. 1989 May;6(3):213-25 PMID: 2560115
  13. Gene splicing by overlap extension: tailor-made genes using the polymerase chain reaction.
    Biotechniques. 1990 May;8(5):528-35 PMID: 2357375
  14. Characterization of the tet(M) determinant of Tn916: evidence for regulation by transcription attenuation.
    Antimicrob Agents Chemother. 1992 Apr;36(4):769-78 PMID: 1323953
  15. Construction of a sequenced Clostridium perfringens-Escherichia coli shuttle plasmid.
    Plasmid. 1992 May;27(3):207-19 PMID: 1513878
  16. Expression of the tetK gene from Staphylococcus aureus in Escherichia coli: comparison of substrate specificities of TetA(B), TetA(C), and TetK efflux proteins.
    Antimicrob Agents Chemother. 1993 Feb;37(2):191-8 PMID: 8452348
  17. Promoter recognition by Escherichia coli RNA polymerase. Effects of single base pair deletions and insertions in the spacer DNA separating the -10 and -35 regions are dependent on spacer DNA sequence.
    Biochemistry. 1993 Jun 22;32(24):6134-40 PMID: 8512922
  18. 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):403-15 PMID: 8170402
  19. Identification and molecular analysis of a locus that regulates extracellular toxin production in Clostridium perfringens.
    Mol Microbiol. 1994 Jun;12(5):761-77 PMID: 8052128
  20. A Clostridium perfringens vector for the selection of promoters.
    Plasmid. 1994 May;31(3):317-9 PMID: 8058826
  21. DNase I footprinting using PCR-generated end-labeled DNA probes.
    Methods Mol Biol. 1995;37:379-91 PMID: 7780518
  22. Molecular genetics of the chloramphenicol-resistance transposon Tn4451 from Clostridium perfringens: the TnpX site-specific recombinase excises a circular transposon molecule.
    Mol Microbiol. 1995 May;16(3):535-51 PMID: 7565113
  23. The closely related ermB-ermAM genes from Clostridium perfringens, Enterococcus faecalis (pAM beta 1), and Streptococcus agalactiae (pIP501) are flanked by variants of a directly repeated sequence.
    Antimicrob Agents Chemother. 1995 Aug;39(8):1830-4 PMID: 7486927
  24. Genetic organization and distribution of tetracycline resistance determinants in Clostridium perfringens.
    Antimicrob Agents Chemother. 1996 Nov;40(11):2500-4 PMID: 8913453
  25. Glutamate residues located within putative transmembrane helices are essential for TetA(P)-mediated tetracycline efflux.
    J Bacteriol. 1997 Nov;179(22):7011-5 PMID: 9371447
  26. Chloramphenicol resistance in Clostridium difficile is encoded on Tn4453 transposons that are closely related to Tn4451 from Clostridium perfringens.
    Antimicrob Agents Chemother. 1998 Jul;42(7):1563-7 PMID: 9660983
  27. Bacillus subtilis tetA(L) gene expression: evidence for regulation by translational reinitiation.
    Mol Microbiol. 1998 Dec;30(5):923-32 PMID: 9988470
  28. Expanded sequence dependence of thermodynamic parameters improves prediction of RNA secondary structure.
    J Mol Biol. 1999 May 21;288(5):911-40 PMID: 10329189
  29. 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):835-58 PMID: 1702475
  30. Molecular genetics and pathogenesis of Clostridium perfringens.
    Microbiol Rev. 1991 Dec;55(4):621-48 PMID: 1779929
  31. Parameters affecting transcription termination by Escherichia coli RNA polymerase. I. Analysis of 13 rho-independent terminators.
    J Mol Biol. 1992 Mar 5;224(1):31-51 PMID: 1372365
  32. Identification of structural and functional domains of the tetracycline efflux protein TetA(P) from Clostridium perfringens.
    Microbiology. 1999 Oct;145 ( Pt 10):2947-55 PMID: 10537217
  33. Changes in the 17 bp spacer in the P(R) promoter of bacteriophage lambda affect steps in open complex formation that precede DNA strand separation.
    J Mol Biol. 2000 Jun 2;299(2):337-49 PMID: 10860742
  34. Transposition of Tn4451 and Tn4453 involves a circular intermediate that forms a promoter for the large resolvase, TnpX.
    Mol Microbiol. 2000 Nov;38(3):588-601 PMID: 11069682
  35. TetZ, a new tetracycline resistance determinant discovered in gram-positive bacteria, shows high homology to gram-negative regulated efflux systems.
    Plasmid. 2000 Nov;44(3):285-91 PMID: 11078655
  36. Induction of pCW3-encoded tetracycline resistance in Clostridium perfringens involves a host-encoded factor.
    Plasmid. 2001 Nov;46(3):229-32 PMID: 11735372
  37. Taxonomy of the Clostridia: ribosomal ribonucleic acid homologies among the species.
    J Gen Microbiol. 1975 Jun;88(2):229-44 PMID: 168308
  38. Isolation and characterization of multiply antibiotic-resistant Clostridum perfringens strains from porcine feces.
    Antimicrob Agents Chemother. 1978 May;13(5):871-80 PMID: 208463
  39. Identification of a transferable tetracycline resistance plasmid (pCW3) from Clostridium perfringens.
    Plasmid. 1978 Sep;1(4):563-70 PMID: 219433
  40. Spacer mutations in the lac ps promoter.
    Proc Natl Acad Sci U S A. 1982 Feb;79(4):1069-72 PMID: 6951162
  41. Compilation and analysis of Escherichia coli promoter DNA sequences.
    Nucleic Acids Res. 1983 Apr 25;11(8):2237-55 PMID: 6344016
  42. Complete nucleotide sequence of pT181, a tetracycline-resistance plasmid from Staphylococcus aureus.
    Plasmid. 1983 Nov;10(3):251-9 PMID: 6657777
  43. Transferable tetracycline resistance in Clostridium perfringens strains of porcine origin.
    Can J Microbiol. 1983 Oct;29(10):1241-6 PMID: 6318939
  44. Molecular analysis of transferable tetracycline resistance plasmids from Clostridium perfringens.
    J Bacteriol. 1985 Feb;161(2):636-40 PMID: 2857166
  45. Characterization in vitro of the effect of spacer length on the activity of Escherichia coli RNA polymerase at the TAC promoter.
    J Biol Chem. 1985 Mar 25;260(6):3529-38 PMID: 3882710
  46. Cloning and analysis of the Clostridium perfringens tetracycline resistance plasmid, pCW3.
    Plasmid. 1985 May;13(3):155-62 PMID: 2860679
  47. Expression of tetracycline resistance in pBR322 derivatives reduces the reproductive fitness of plasmid-containing Escherichia coli.
    Gene. 1985;39(2-3):173-80 PMID: 3005111
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.183.24.7110-7119.2001
Published
2001-12-00
Pages
7110-9
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC95559
Subset
IM
External Links
PubMed source DOI: 10.1128/JB.183.24.7110-7119.2001 PMC Full Text View journal details
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