1887

Abstract

Resistance of strain HB251 to the newer -lactam antibiotics, in particular ceftazidime and aztreonam, results from production of the extended-spectrum -lactamase TEM-6. The corresponding structural gene, , and its promoter region were amplified by the polymerase chain reaction. Analysis of the sequence of the amplification product showed that differed by two nucleotide substitutions from , the gene encoding TEM-1 penicillinase in plasmid pBR322. The mutations led to the substitution of a lysine for a glutamic acid at position 102 and of a histidine for an arginine at position 162 of the unprocessed TEM-1 protein. The presence of a 116 bp DNA insert upstream from resulted in the creation of hybrid promoter P in which the –10 region was that of TEM-1 promoter P whereas the – 35 canonical sequence TTGACA was provided by the right end of the insert. P was found to be 10 times more active than P and to confer higher levels of antibiotic resistance upon the host. Analysis of the sequence of the insert indicated that the 116 bp fragment is related to insertion sequence IS but differs from it by three internal deletions that removed regions encoding the transposase. The distribution of the IS-like element in clinical isolates of was studied by the polymerase chain reaction and by DNA-DNA hybridization. The element appeared to be widespread and was detected in strains producing TEM-6 or other TEM variants.

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1991-12-01
2021-05-11
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References

  1. Barthélémy M., Guionie M., Labia R. 1978; β-Lactamases : determination of their isoelectric points. Antimicrobial Agents and Chemotherapy 13:695–698
    [Google Scholar]
  2. Barthélémy M., Péduzzi J., Ben Yaghlane H., Labia R. 1988; Single amino acid substitution between SHV-1 β-lactamase and cefotaxime-hydrolyzing SHV-2 enzyme. FEBS Letters 231:217–220
    [Google Scholar]
  3. Bauernfeind A., Hörl G. 1987; Novel R-factor borne β-lactamase of Escherichia coli conferring resistance to cephalosporins. Infection 15:257–259
    [Google Scholar]
  4. Bauernfeind A., Ključar S. V., Pritzbuer E., Medeiros A. 1989; Epidemiology of resistance due to TEM-6 β-lactamase in enterobacteriaceae in intensive care patients. Abstracts of the 4th European Congress of Clinical Microbiology abstract no. 1125
    [Google Scholar]
  5. Bolivar F., Rodriguez R. L., Betlach M. C., Boyer H. W. 1977; Construction and characterization of new cloning vehicles. I. Ampicillin-resistant derivatives of the plasmid pMB9. Gene 2:75–93
    [Google Scholar]
  6. Boyer H. W., Roulland-Dussoix D. 1969; A complementation analysis of the restriction and modification of DNA in Escherichia coli . Journal of Molecular Biology 41:459–472
    [Google Scholar]
  7. Brisson-Noël A., Arthur M., Courvalin P. 1988; Evidence for natural gene transfer from gram-positive cocci to Escherichia coli . Journal of Bacteriology 170:1739–1745
    [Google Scholar]
  8. Brosius J. 1984; Plasmid vector for the selection of promoters. Gene 27:151–160
    [Google Scholar]
  9. Chabbert Y. A., Derlot E. 1983; Comparative activity of cefotetan on Escherichia coli K12 possessing plasmid-mediated β-lactamases. Journal of Antimicrobial Chemotherapy 11:159–167
    [Google Scholar]
  10. Chen S. T., Clowes R. C. 1984; Two improved promoter sequences for the β-lactamase expression arising from a single base-pair substitution. Nucleic Acids Research 12:3219–3234
    [Google Scholar]
  11. Collatz E., Tran Van Nhieu G., Billot-Klein D., , Williamson R., Gutmann L. 1989; Substitution of serine for arginine in position 162 of TEM-type β-lactamases extends the substrate profile of mutant enzymes, TEM-7 and TEM-101, to ceftazidime and aztreonam. Gene 78:349–354
    [Google Scholar]
  12. Grindley N. D., Reed R. R. 1985; Transpositional recombination in prokaryotes. Annual Review of Biochemistry 54:863–896
    [Google Scholar]
  13. Grunstein M., Hogness D. 1975; Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proceedings of the National Academy of Sciences of the United States of America 723961–3965
    [Google Scholar]
  14. Hawley D. K., McClure D. W. 1983; Compilation and analysis of Escherichia coli promoter DNA sequence. Nucleic Acids Research 11:2237–2255
    [Google Scholar]
  15. Herzberg O., Moult J. 1987; Bacterial resistance to β-lactam antibiotics: crystal structure of β-lactamase from Staphylococcus aureus PCI at 2-5Å resolution. Science 236:694–701
    [Google Scholar]
  16. Johnson D. A., Sportsman J., Elder J. 1984; Improved technique utilizing nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Analysis Techniques 1:3–8
    [Google Scholar]
  17. Kliebe C., Nies B. A., Meyer J. F., Tolxdorff-Neutzling R. M., Wiedemann B. 1985; Evolution of plasmid-coded resistance to broad-spectrum cephalosporins. Antimicrobial Agents and Chemotherapy 28:302–307
    [Google Scholar]
  18. Knothe H., Shah P., Krcméry V., Antal M., Mitsuhashi S. 1983; Transferable resistance to cefotaxime, cefoxitin, cefamandole and cefuroxime in clinical isolates of Klebsiella pneumoniae and Serratia marcescens . Infection 11:315–317
    [Google Scholar]
  19. Labigne-Roussel A., , Gerbaud G., Courvalin P. 1981; Translocation of sequences encoding antibiotic resistance from the chromosome to a receptor plasmid in Salmonella ordonez . Molecular and General Genetics 182:390–408
    [Google Scholar]
  20. Lupski J. R., Ruiz A. A., Godson G. N. 1984; Promotion, termination, and anti-termination in the rpsU-dnaG-rpoD macromo-lecular synthesis operon of E. coli K-12. Molecular and General Genetics 195:391–401
    [Google Scholar]
  21. Mabilat C., Courvalin P. 1990; Development of ‘oligotyping’ for characterization and molecular epidemiology of TEM β-lactamases in Enterobacteriaceae . Antimicrobial Agents and Chemotherapy 34:2210–2216
    [Google Scholar]
  22. Mabilat C., Goussard S., Sougakoff W., Spencer R. C., Courvalin P. 1990; Direct sequencing of the amplified structural gene and promoter for the extended-broad-spectrum β-lactamase TEM-9 (RHH-1) of Klebsiella pneumoniae . Plasmid 23:27–34
    [Google Scholar]
  23. Maniatis T., Fritsch E. F., Sambrook J. 1982; Molecular Cloning. A Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  24. Matthew M., Hedges R. W. 1976; Analytical isoelectric focusing of R factor-determined β-lactamases : correlation with plasmid compatibility. Journal of Bacteriology 125:713–718
    [Google Scholar]
  25. Medeiros A. A. 1989; Plasmid-determined β-lactamases. Microbial Resistance to Drugs101–127 Bryan L. E. Berlin: Springer-Verlag;
    [Google Scholar]
  26. Moran C. P., Lang N., LeGrice S. F., Lee G., Stephens M., Sonenshein A. L., Pero J., Losick R. 1982; Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis . Molecular and General Genetics 186:339–346
    [Google Scholar]
  27. Ohtsubo E., Ohtsubo H. 1978; Nucleotide sequence of an insertion element ISI . Proceedings of the National Academy of Sciences of the United States of America 75615–619
    [Google Scholar]
  28. Paul G. C., Gerbaud G., Bure A., Philippon A. M., Pangon B., Courvalin P. 1989; TEM-4, a new plasmid-mediated β-lactamase that hydrolyzes broad-spectrum cephalosporins in a clinical isolate of Escherichia coli . Antimicrobial Agents and Chemotherapy 33:1958–1963
    [Google Scholar]
  29. Petit A., Sirot D. L., Chanal C. M., Sirot J. L., Labia R., Gerbaud G., Cluzel R. A. 1988; Novel plasmid-mediated β-lactamase in clinical isolates of Klebsiella pneumoniae more resistant to ceftazidime than to other broad-spectrum cephalosporins. Antimicrobial Agents and Chemotherapy 32:626–630
    [Google Scholar]
  30. Philippon A., Labia R., Jacoby G. 1989; Extended-spectrum β-lactamases. Antimicrobial Agents and Chemotherapy 33:1131–1136
    [Google Scholar]
  31. Prentki P., Teter B., Chandler M., Galas D. J. 1986; Functional promoters created by the insertion of transposable element ISI . Journal of Molecular Biology 191:383–393
    [Google Scholar]
  32. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. 1988; Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–494
    [Google Scholar]
  33. Sirot D., Sirot J., Labia R., Morand R., Courvalin P., Darfeuille-Michaud A., , Perroux R., Cluzel R. 1987; Transferable resistance to third-generation cephalosporins in clinical isolates of Klebsiella pneumoniae: identification of CTX-1, a novel β-lactamase. Journal of Antimicrobial Chemotherapy 20:323–334
    [Google Scholar]
  34. Sougakoff W., Goussard S., Courvalin P. 1988a; Plasmid-mediated resistance to third-generation cephalosporins caused by point mutations in TEM-type penicillinase genes. Reviews of Infectious Diseases 10:879–884
    [Google Scholar]
  35. Sougakoff W., Goussard S., Courvalin P. 1988b; The TEM-3 β-lactamase, which hydrolyzes broad-spectrum cephalosporins, is derived from the TEM-2 penicillinase by two amino acid substitutions. FEMS Letters 56:343–348
    [Google Scholar]
  36. Sougakoff W., Petit A., Goussard S., Sirot D., Bure A., Courvalin P. 1989; Characterization of the plasmid genes blaT-4 and blaT-5 which encode the broad-spectrum β-lactamases TEM-4 and TEM-5 in Enterobacteriaceae . Gene 78:339–348
    [Google Scholar]
  37. Southern E. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98:503–517
    [Google Scholar]
  38. Spencer R. C., Wheat P. F., Winstanley P. G., Cox D. M., Plested S. J. 1987; Novel β-lactamase in a clinical isolate of Klebsiella pneumoniae conferring unusual resistance to β-lactam antibiotics. Journal of Antimicrobial Chemotherapy 20:919–927
    [Google Scholar]
  39. Sutcliffe J. G. 1978; Nucleotide sequence of the ampicillin resistance gene of Escherichia coli plasmid pBR322. Proceedings of the National Academy of Sciences of the United States of America 753737–3741
    [Google Scholar]
  40. Tabor S., Richardson C. C. 1987; DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proceedings of the National Academy of Sciences of the United States of America 844767–4771
    [Google Scholar]
  41. Wilbur W. J., Lipman D. J. 1983; Rapid similarity searches of nucleic acid and protein data banks. Proceedings of the National Academy of Sciences of the United States of America 80726–730
    [Google Scholar]
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