1887

Abstract

subsp. strain ATTn10 produces the -lactam antibiotic 1-carbapen-2-em-3-carboxylic acid (carbapenem) by expressing the operon. Mutants exhibiting increased carbapenem gene transcription were positively selected using an engineered strain with a functional -lactamase translational fusion in , the last gene of the operon. However, spontaneous ampicillin-resistant mutants were isolated even when transcription of  : :  was blocked by a strongly polar mutation in . The mechanism of resistance was shown to be due to cryptic IS elements transposing upstream of  : : , thereby providing new promoters enabling  : :  transcription. Southern blots showed that IS was present in multicopy in ATTn10. In addition, a Tn genetic remnant was discovered. The results offer insights into the genetic archaeology of strain ATTn10 and highlight the powerful impacts of cryptic IS elements in bacterial adaptive evolution.

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2006-04-01
2019-11-12
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References

  1. Albertini, A. M., Hofer, M., Calos, M. P. & Miller, J. H. ( 1982; ). On the formation of spontaneous deletions: the importance of short sequence homologies in the generation of large deletions. Cell 29, 319–328.[CrossRef]
    [Google Scholar]
  2. Altschul, S. F., Gish, W., Miller, W., Myers, E. W. & Lipman, D. J. ( 1990; ). Basic local alignment search tool. J Mol Biol 215, 403–410.[CrossRef]
    [Google Scholar]
  3. Bainton, N. J., Stead, P., Chhabra, S. R., Bycroft, B. W., Salmond, G. P. C., Stewart, G. S. A. B. & Williams, P. ( 1992; ). N-(3-oxohexanoyl)-l-homoserine lactone regulates carbapenem antibiotic production in Erwinia carotovora. Biochem J 288, 997–1004.
    [Google Scholar]
  4. Balbinder, E. ( 2001; ). Stationary phase deletions in Escherichia coli. I – evidence for a new deletion pathway. Mutat Res 479, 19–36.[CrossRef]
    [Google Scholar]
  5. Bender, J. & Kleckner, N. ( 1986; ). Genetic evidence that Tn10 transposes by a nonreplicative mechanism. Cell 45, 801–815.[CrossRef]
    [Google Scholar]
  6. Bender, J., Kuo, J. & Kleckner, N. ( 1991; ). Genetic evidence against intramolecular rejoining of the donor DNA molecule following IS10 transposition. Genetics 128, 687–694.
    [Google Scholar]
  7. Bochner, B. R., Huang, H., Schieven, G. L. & Ames, B. N. ( 1980; ). Positive selection for loss of tetracycline resistance. J Bacteriol 143, 926–933.
    [Google Scholar]
  8. Bogosian, G., Bilyeu, K. & O'Neil, J. P. ( 1993; ). Genome rearrangements by residual IS10 elements in strains of Escherichia coli K-12 which had undergone Tn10 mutagenesis and fusaric acid selection. Gene 133, 17–22.[CrossRef]
    [Google Scholar]
  9. Chalmers, R. M. & Kleckner, N. ( 1996; ). IS10/Tn10 transposition efficiently accommodates diverse transposon configurations. EMBO J 15, 5112–5122.
    [Google Scholar]
  10. Chalmers, R., Sewitz, S., Lipkow, K. & Crellin, P. ( 2000; ). Complete nucleotide sequence of Tn10. J Bacteriol 182, 2970–2972.[CrossRef]
    [Google Scholar]
  11. Ciampi, M. S., Schmid, M. B. & Roth, J. R. ( 1982; ). Transposon Tn10 provides a promoter for transcription of adjacent sequences. Proc Natl Acad Sci U S A 79, 5016–5020.[CrossRef]
    [Google Scholar]
  12. Coulthurst, S. J., Barnard, A. M. L. & Salmond, G. P. C. ( 2005; ). Regulation and biosynthesis of carbapenem antibiotics in bacteria. Nat Rev Microbiol 3, 295–306.[CrossRef]
    [Google Scholar]
  13. Eichenbaum, Z. & Livneh, Z. ( 1998; ). UV light induces IS10 transposition in Escherichia coli. Genetics 149, 1173–1181.
    [Google Scholar]
  14. Farabaugh, P. J., Schmeissner, U., Hofer, M. & Miller, J. H. ( 1978; ). Genetic studies of the lac repressor. VII. On the molecular nature of spontaneous hotspots in the lacI gene of Escherichia coli. J Mol Biol 126, 847–857.[CrossRef]
    [Google Scholar]
  15. Grinter, N. J. ( 1983; ). A broad-host-range cloning vector transposable to various replicons. Gene 21, 133–143.[CrossRef]
    [Google Scholar]
  16. Halling, S. M. & Kleckner, N. ( 1982; ). A symmetrical six-base-pair target site sequence determines Tn10 insertion specificity. Cell 28, 155–163.[CrossRef]
    [Google Scholar]
  17. Herrero, M., De Lorenzo, V. & Timmis, K. N. ( 1990; ). Transposon vectors containing non-antibiotic resistance selection markers for cloning and stable chromosomal insertion of foreign genes in Gram-negative bacteria. J Bacteriol 172, 6557–6567.
    [Google Scholar]
  18. Jellen-Ritter, A. S. & Kern, W. V. ( 2001; ). Enhanced expression of the multidrug efflux pumps AcrAB and AcrEF associated with insertion element transposition in Escherichia coli mutants selected with a fluoroquinolone. Antimicrob Agents Chemother 45, 1467–1472.[CrossRef]
    [Google Scholar]
  19. Kaniga, K., Delor, I. & Cornelis, G. R. ( 1991; ). A wide-host-range suicide vector for improving reverse genetics in Gram-negative bacteria: inactivation of the blaA gene of Yersinia enterocolitica. Gene 109, 137–141.[CrossRef]
    [Google Scholar]
  20. Kleckner, N., Chalmers, R. M., Kwon, D., Sakai, J. & Bolland, S. ( 1995; ). Tn10 and IS10 transposition and chromosome rearrangements: mechanism and regulation in vivo and in vitro. Curr Top Microbiol Immunol 204, 49–82.
    [Google Scholar]
  21. Kobayashi, K., Tsukagoshi, N. & Aono, R. ( 2001; ). Suppression of hypersensitivity of Escherichia coli acrB mutant to organic solvents by integrational activation of the acrEF operon with the IS1 or IS2 element. J Bacteriol 183, 2646–2653.[CrossRef]
    [Google Scholar]
  22. Marvo, S. L., King, S. R. & Jaskunas, S. R. ( 1983; ). Role of short regions of homology in intermolecular illegitimate recombination events. Proc Natl Acad Sci U S A 80, 2452–2456.[CrossRef]
    [Google Scholar]
  23. Matsutani, S. ( 1991; ). Multiple copies of IS10 in the Enterobacter cloacae MD36 chromosome. J Bacteriol 173, 7802–7809.
    [Google Scholar]
  24. McGowan, S. J., Sebaihia, M., Porter, L. E., Stewart, G. S. A. B., Williams, P., Bycroft, B. W. & Salmond, G. P. C. ( 1996; ). Analysis of bacterial carbapenem antibiotic production genes reveals a novel β-lactam biosynthesis pathway. Mol Microbiol 22, 415–426.[CrossRef]
    [Google Scholar]
  25. McGowan, S. J., Sebaihia, M., O'Leary, S., Hardie, K. R., Williams, P., Stewart, G. S. A. B., Bycroft, B. W. & Salmond, G. P. C. ( 1997; ). Analysis of the carbapenem gene cluster of Erwinia carotovora: definition of the antibiotic biosynthetic genes and evidence for a novel β-lactam resistance mechanism. Mol Microbiol 26, 545–556.[CrossRef]
    [Google Scholar]
  26. McGowan, S. J., Barnard, A. M. L., Bosgelmez, G. & 7 other authors ( 2005; ). Carbapenem antibiotic biosynthesis in Erwinia carotovora is regulated by physiological and genetic factors modulating the quorum sensing-dependent control pathway. Mol Microbiol 55, 526–545.
    [Google Scholar]
  27. Miller, J. H. ( 1972; ). Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  28. Miller, C., Thomsen, L. E., Gaggero, C., Mosseri, R., Ingmer, H. & Cohen, S. N. ( 2004; ). SOS response induction by β-lactams and bacterial defense against antibiotic lethality. Science 305, 1629–1631.[CrossRef]
    [Google Scholar]
  29. Olliver, A., Valle, M., Chaslus-Dancla, E. & Cloeckaert, A. ( 2005; ). Overexpression of the multidrug efflux operon acrEF by insertional activation with IS1 or IS10 elements in Salmonella enterica serovar Typhimurium DT204 acrB mutants selected with fluoroquinolones. Antimicrob Agents Chemother 49, 289–301.[CrossRef]
    [Google Scholar]
  30. Prentki, P. & Krisch, H. M. ( 1984; ). In vitro insertional mutagenesis with a selectable DNA fragment. Gene 29, 303–313.[CrossRef]
    [Google Scholar]
  31. Reading, C. & Cole, M. ( 1977; ). Clavulanic acid: a beta-lactamase-inhibiting beta-lactam from Streptomyces clavuligerus. Antimicrob Agents Chemother 11, 852–857.[CrossRef]
    [Google Scholar]
  32. Roberts, D., Hoopes, B. C., McClure, W. R. & Kleckner, N. ( 1985; ). IS10 transposition is regulated by DNA adenine methylation. Cell 43, 117–130.[CrossRef]
    [Google Scholar]
  33. Roberts, D. E., Ascherman, D. & Kleckner, N. ( 1991; ). IS10 promotes adjacent deletions at low frequency. Genetics 128, 37–43.
    [Google Scholar]
  34. Sambrook, J., Fritsch, E. F. & Maniatis, T. ( 1989; ). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  35. Simons, R. W. & Kleckner, N. ( 1983; ). Translational control of IS10 transposition. Cell 34, 683–691.[CrossRef]
    [Google Scholar]
  36. Simons, R. W., Hoopes, B. C., McClure, W. R. & Kleckner, N. ( 1983; ). Three promoters near the termini of IS10: pIN, pOUT, and pIII. Cell 34, 673–682.[CrossRef]
    [Google Scholar]
  37. Singer, B. S. & Westlye, J. ( 1988; ). Deletion formation in bacteriophage T4. J Mol Biol 202, 233–243.[CrossRef]
    [Google Scholar]
  38. Toth, I., Perombelon, M. C. M. & Salmond, G. P. C. ( 1993; ). Bacteriophage ϕKP mediated generalised transduction in Erwinia carotovora subsp. carotovora. J Gen Microbiol 139, 2705–2709.[CrossRef]
    [Google Scholar]
  39. Uematsu, N., Matsuoka, C., Agemizu, Y., Nagoshi, E. & Yamamoto, K. ( 1999; ). Asymmetric crossing over in the spontaneous formation of large deletions in the tonB-trp region of the Escherichia coli K-12 chromosome. Mol Gen Genet 261, 523–529.[CrossRef]
    [Google Scholar]
  40. Welch, M., Todd, D. E., Whitehead, N. A., McGowan, S. J., Bycroft, B. W. & Salmond, G. P. C. ( 2000; ). N-acyl homoserine lactone binding to the CarR receptor determines quorum-sensing specificity in Erwinia. EMBO J 19, 631–641.[CrossRef]
    [Google Scholar]
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