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Journal of Bacteriology logoLink to Journal of Bacteriology
. 1987 May;169(5):1923–1928. doi: 10.1128/jb.169.5.1923-1928.1987

Inactivation of the ampD gene causes semiconstitutive overproduction of the inducible Citrobacter freundii beta-lactamase.

F Lindberg, S Lindquist, S Normark
PMCID: PMC212046  PMID: 3032901

Abstract

In Citrobacter freundii and Enterobacter cloacae, synthesis of AmpC beta-lactamase is inducible by the addition of beta-lactams to the growth medium. Spontaneous mutants that constitutively overproduce the enzyme occur at a high frequency. When the C. freundii ampC beta-lactamase gene is cloned into Escherichia coli together with the regulatory gene ampR, beta-lactamase expression from the clone is inducible. Spontaneous cefotaxime-resistant mutants were selected from an E. coli strain carrying the cloned C. freundii ampC and ampR genes on a plasmid. Virtually all isolates had chromosomal mutations leading to semiconstitutive overproduction of beta-lactamase. The mutation ampD2 in one such mutant was caused by an IS1 insertion into the hitherto unknown ampD gene, located between nadC and aroP at minute 2.4 on the E. coli chromosome. The wild-type ampD allele cloned on a plasmid could fully trans-complement beta-lactamase-overproducing mutants of both E. coli and C. freundii, restoring the wild-type phenotype of highly inducible enzyme synthesis. This indicates that these E. coli and C. freundii mutants have their lesions in ampD. We hypothesize that induction of beta-lactamase synthesis is caused by blocking of the AmpD function by the beta-lactam inducer and that this leads directly or indirectly to an AmpR-mediated stimulation of ampC expression.

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Selected References

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  1. BERTANI G. Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J Bacteriol. 1951 Sep;62(3):293–300. doi: 10.1128/jb.62.3.293-300.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bergström S., Lindberg F. P., Olsson O., Normark S. Comparison of the overlapping frd and ampC operons of Escherichia coli with the corresponding DNA sequences in other gram-negative bacteria. J Bacteriol. 1983 Sep;155(3):1297–1305. doi: 10.1128/jb.155.3.1297-1305.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Björk G. R., Olsén A. A method for isolation of Escherichia coli mutants with aberrant RNA methylation using translocatable drug resistance elements. Acta Chem Scand B. 1979;33(8):591–593. doi: 10.3891/acta.chem.scand.33b-0591. [DOI] [PubMed] [Google Scholar]
  5. Burman L. G., Park J. T., Lindström E. B., Boman H. G. Resistance of Escherichia coli to penicillins: identification of the structural gene for the chromosomal penicillinase. J Bacteriol. 1973 Oct;116(1):123–130. doi: 10.1128/jb.116.1.123-130.1973. [DOI] [PMC free article] [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. Chye M. L., Guest J. R., Pittard J. Cloning of the aroP gene and identification of its product in Escherichia coli K-12. J Bacteriol. 1986 Aug;167(2):749–753. doi: 10.1128/jb.167.2.749-753.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gootz T. D., Jackson D. B., Sherris J. C. Development of resistance to cephalosporins in clinical strains of Citrobacter spp. Antimicrob Agents Chemother. 1984 May;25(5):591–595. doi: 10.1128/aac.25.5.591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Guest J. R., Roberts R. E., Stephens P. E. Hybrid plasmids containing the pyruvate dehydrogenase complex genes and gene-DNA relationships in the 2 to 3 minute region of the Escherichia coli chromosome. J Gen Microbiol. 1983 Mar;129(3):671–680. doi: 10.1099/00221287-129-3-671. [DOI] [PubMed] [Google Scholar]
  10. Guest J. R., Stephens P. E. Molecular cloning of the pyruvate dehydrogenase complex genes of Escherichia coli. J Gen Microbiol. 1980 Dec;121(2):277–292. doi: 10.1099/00221287-121-2-277. [DOI] [PubMed] [Google Scholar]
  11. Hall M. N., Silhavy T. J. Genetic analysis of the ompB locus in Escherichia coli K-12. J Mol Biol. 1981 Sep 5;151(1):1–15. doi: 10.1016/0022-2836(81)90218-7. [DOI] [PubMed] [Google Scholar]
  12. Jaurin B., Grundström T. ampC cephalosporinase of Escherichia coli K-12 has a different evolutionary origin from that of beta-lactamases of the penicillinase type. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4897–4901. doi: 10.1073/pnas.78.8.4897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Joris B., De Meester F., Galleni M., Reckinger G., Coyette J., Frere J. M., Van Beeumen J. The beta-lactamase of Enterobacter cloacae P99. Chemical properties, N-terminal sequence and interaction with 6 beta-halogenopenicillanates. Biochem J. 1985 May 15;228(1):241–248. doi: 10.1042/bj2280241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. Lindberg F., Normark S. Common mechanism of ampC beta-lactamase induction in enterobacteria: regulation of the cloned Enterobacter cloacae P99 beta-lactamase gene. J Bacteriol. 1987 Feb;169(2):758–763. doi: 10.1128/jb.169.2.758-763.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lindberg F., Normark S. Sequence of the Citrobacter freundii OS60 chromosomal ampC beta-lactamase gene. Eur J Biochem. 1986 May 2;156(3):441–445. doi: 10.1111/j.1432-1033.1986.tb09601.x. [DOI] [PubMed] [Google Scholar]
  18. Lindberg F., Westman L., Normark S. Regulatory components in Citrobacter freundii ampC beta-lactamase induction. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4620–4624. doi: 10.1073/pnas.82.14.4620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Low B. Rapid mapping of conditional and auxotrophic mutations in Escherichia coli K-12. J Bacteriol. 1973 Feb;113(2):798–812. doi: 10.1128/jb.113.2.798-812.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Low K. B. Escherichia coli K-12 F-prime factors, old and new. Bacteriol Rev. 1972 Dec;36(4):587–607. doi: 10.1128/br.36.4.587-607.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mandel M., Higa A. Calcium-dependent bacteriophage DNA infection. J Mol Biol. 1970 Oct 14;53(1):159–162. doi: 10.1016/0022-2836(70)90051-3. [DOI] [PubMed] [Google Scholar]
  22. Normark S., Burman L. G. Resistance of Escherichia coli to penicillins: fine-structure mapping and dominance of chromosomal beta-lactamase mutations. J Bacteriol. 1977 Oct;132(1):1–7. doi: 10.1128/jb.132.1.1-7.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Normark S. Genetics of a chain-forming mutant of Escherichia coli. Transduction and dominance of the envA gene mediating increased penetration to some antibacterial agents. Genet Res. 1970 Aug;16(1):63–78. doi: 10.1017/s0016672300002287. [DOI] [PubMed] [Google Scholar]
  24. Ohtsubo H., Ohtsubo E. Nucleotide sequence of an insertion element, IS1. Proc Natl Acad Sci U S A. 1978 Feb;75(2):615–619. doi: 10.1073/pnas.75.2.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Olsson O., Bergström S., Normark S. Identification of a novel ampC beta-lactamase promoter in a clinical isolate of Escherichia coli. EMBO J. 1982;1(11):1411–1416. doi: 10.1002/j.1460-2075.1982.tb01331.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sanders C. C., Sanders W. E., Jr Emergence of resistance during therapy with the newer beta-lactam antibiotics: role of inducible beta-lactamases and implications for the future. Rev Infect Dis. 1983 Jul-Aug;5(4):639–648. doi: 10.1093/clinids/5.4.639. [DOI] [PubMed] [Google Scholar]
  27. Shattuck-Eidens D. M., Kadner R. J. Exogenous induction of the Escherichia coli hexose phosphate transport system defined by uhp-lac operon fusions. J Bacteriol. 1981 Oct;148(1):203–209. doi: 10.1128/jb.148.1.203-209.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Shattuck-Eidens D. M., Kadner R. J. Molecular cloning of the uhp region and evidence for a positive activator for expression of the hexose phosphate transport system of Escherichia coli. J Bacteriol. 1983 Sep;155(3):1062–1070. doi: 10.1128/jb.155.3.1062-1070.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stoker N. G., Fairweather N. F., Spratt B. G. Versatile low-copy-number plasmid vectors for cloning in Escherichia coli. Gene. 1982 Jun;18(3):335–341. doi: 10.1016/0378-1119(82)90172-x. [DOI] [PubMed] [Google Scholar]
  30. Sykes R. B., Matthew M. The beta-lactamases of gram-negative bacteria and their role in resistance to beta-lactam antibiotics. J Antimicrob Chemother. 1976 Jun;2(2):115–157. doi: 10.1093/jac/2.2.115. [DOI] [PubMed] [Google Scholar]
  31. Tritz G. J., Matney T. S., Chandler J. L., Gholson R. K. Chromosomal location of the C gene involved in the biosynthesis of nicotinamide adenine dinucleotide in Escherichia coli K-12. J Bacteriol. 1970 Oct;104(1):45–49. doi: 10.1128/jb.104.1.45-49.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  33. Waxman D. J., Strominger J. L. Penicillin-binding proteins and the mechanism of action of beta-lactam antibiotics. Annu Rev Biochem. 1983;52:825–869. doi: 10.1146/annurev.bi.52.070183.004141. [DOI] [PubMed] [Google Scholar]

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