Skip to main content
PMC home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1996 Sep;40(9):2029–2033. doi: 10.1128/aac.40.9.2029

A rob-like gene of Enterobacter cloacae affecting porin synthesis and susceptibility to multiple antibiotics.

E H Lee 1, E Collatz 1, I Podglajen 1, L Gutmann 1
PMCID: PMC163467  PMID: 8878575

Abstract

A chromosomal gene of Enterobacter cloacae affecting the synthesis of major outer membrane proteins in E. cloacae and Escherichia coli was cloned by using selection for resistance to cefoxitin in E. coli. The presence of the gene, when plasmid-borne, led to a decrease in the amount of porin F in E. cloacae and the amount of OmpF in E. coli and caused 2- to 32-fold increases in the MICs of chloramphenicol, tetracycline, quinolones, and beta-lactam antibiotics. The gene encoded a 33-kDa protein, similar (83% identity) to the protein Rob involved in the initiation of DNA replication in E. coli, which was called RobA(EC1) by analogy. RobA from E. cloacae was found to inhibit ompF expression at the posttranscriptional level via activation of micF, a gene also apparently present in E. cloacae, as detected by PCR. As with its homolog from E. coli, RobA(EC1) is related to the XylS-AraC class of positive transcriptional regulators, along with MarA and SoxS, which also cause a micF-mediated decrease in the level of ampF expression.

Full Text

The Full Text of this article is available as a PDF (320.9 KB).

Selected References

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

  1. Andersen J., Forst S. A., Zhao K., Inouye M., Delihas N. The function of micF RNA. micF RNA is a major factor in the thermal regulation of OmpF protein in Escherichia coli. J Biol Chem. 1989 Oct 25;264(30):17961–17970. [PubMed] [Google Scholar]
  2. Ariza R. R., Cohen S. P., Bachhawat N., Levy S. B., Demple B. Repressor mutations in the marRAB operon that activate oxidative stress genes and multiple antibiotic resistance in Escherichia coli. J Bacteriol. 1994 Jan;176(1):143–148. doi: 10.1128/jb.176.1.143-148.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ariza R. R., Li Z., Ringstad N., Demple B. Activation of multiple antibiotic resistance and binding of stress-inducible promoters by Escherichia coli Rob protein. J Bacteriol. 1995 Apr;177(7):1655–1661. doi: 10.1128/jb.177.7.1655-1661.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chou J. H., Greenberg J. T., Demple B. Posttranscriptional repression of Escherichia coli OmpF protein in response to redox stress: positive control of the micF antisense RNA by the soxRS locus. J Bacteriol. 1993 Feb;175(4):1026–1031. doi: 10.1128/jb.175.4.1026-1031.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen S. P., Hächler H., Levy S. B. Genetic and functional analysis of the multiple antibiotic resistance (mar) locus in Escherichia coli. J Bacteriol. 1993 Mar;175(5):1484–1492. doi: 10.1128/jb.175.5.1484-1492.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cohen S. P., McMurry L. M., Levy S. B. marA locus causes decreased expression of OmpF porin in multiple-antibiotic-resistant (Mar) mutants of Escherichia coli. J Bacteriol. 1988 Dec;170(12):5416–5422. doi: 10.1128/jb.170.12.5416-5422.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Coyer J., Andersen J., Forst S. A., Inouye M., Delihas N. micF RNA in ompB mutants of Escherichia coli: different pathways regulate micF RNA levels in response to osmolarity and temperature change. J Bacteriol. 1990 Aug;172(8):4143–4150. doi: 10.1128/jb.172.8.4143-4150.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Decad G. M., Nikaido H. Outer membrane of gram-negative bacteria. XII. Molecular-sieving function of cell wall. J Bacteriol. 1976 Oct;128(1):325–336. doi: 10.1128/jb.128.1.325-336.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Esterling L., Delihas N. The regulatory RNA gene micF is present in several species of gram-negative bacteria and is phylogenetically conserved. Mol Microbiol. 1994 May;12(4):639–646. doi: 10.1111/j.1365-2958.1994.tb01051.x. [DOI] [PubMed] [Google Scholar]
  10. Gallegos M. T., Michán C., Ramos J. L. The XylS/AraC family of regulators. Nucleic Acids Res. 1993 Feb 25;21(4):807–810. doi: 10.1093/nar/21.4.807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gambino L., Gracheck S. J., Miller P. F. Overexpression of the MarA positive regulator is sufficient to confer multiple antibiotic resistance in Escherichia coli. J Bacteriol. 1993 May;175(10):2888–2894. doi: 10.1128/jb.175.10.2888-2894.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. George A. M., Hall R. M., Stokes H. W. Multidrug resistance in Klebsiella pneumoniae: a novel gene, ramA, confers a multidrug resistance phenotype in Escherichia coli. Microbiology. 1995 Aug;141(Pt 8):1909–1920. doi: 10.1099/13500872-141-8-1909. [DOI] [PubMed] [Google Scholar]
  13. Hall M. N., Silhavy T. J. The ompB locus and the regulation of the major outer membrane porin proteins of Escherichia coli K12. J Mol Biol. 1981 Feb 15;146(1):23–43. doi: 10.1016/0022-2836(81)90364-8. [DOI] [PubMed] [Google Scholar]
  14. Hall M. N., Silhavy T. J. Transcriptional regulation of Escherichia coli K-12 major outer membrane protein 1b. J Bacteriol. 1979 Nov;140(2):342–350. doi: 10.1128/jb.140.2.342-350.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hancock R. E. Role of porins in outer membrane permeability. J Bacteriol. 1987 Mar;169(3):929–933. doi: 10.1128/jb.169.3.929-933.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Harder K. J., Nikaido H., Matsuhashi M. Mutants of Escherichia coli that are resistant to certain beta-lactam compounds lack the ompF porin. Antimicrob Agents Chemother. 1981 Oct;20(4):549–552. doi: 10.1128/aac.20.4.549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hawley D. K., McClure W. R. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. doi: 10.1093/nar/11.8.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hooper D. C., Wolfson J. S., Souza K. S., Tung C., McHugh G. L., Swartz M. N. Genetic and biochemical characterization of norfloxacin resistance in Escherichia coli. Antimicrob Agents Chemother. 1986 Apr;29(4):639–644. doi: 10.1128/aac.29.4.639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ingham C., Buechner M., Adler J. Effect of outer membrane permeability on chemotaxis in Escherichia coli. J Bacteriol. 1990 Jul;172(7):3577–3583. doi: 10.1128/jb.172.7.3577-3583.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ishida H., Fuziwara H., Kaibori Y., Horiuchi T., Sato K., Osada Y. Cloning of multidrug resistance gene pqrA from Proteus vulgaris. Antimicrob Agents Chemother. 1995 Feb;39(2):453–457. doi: 10.1128/aac.39.2.453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Komatsu T., Ohta M., Kido N., Arakawa Y., Ito H., Mizuno T., Kato N. Molecular characterization of an Enterobacter cloacae gene (romA) which pleiotropically inhibits the expression of Escherichia coli outer membrane proteins. J Bacteriol. 1990 Jul;172(7):4082–4089. doi: 10.1128/jb.172.7.4082-4089.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lee E. H., Collatz E., Trias J., Gutmann L. Diffusion of beta-lactam antibiotics into proteoliposomes reconstituted with outer membranes of isogenic imipenem-susceptible and -resistant strains of Enterobacter cloacae. J Gen Microbiol. 1992 Nov;138(11):2347–2351. doi: 10.1099/00221287-138-11-2347. [DOI] [PubMed] [Google Scholar]
  23. Lee E. H., Nicolas M. H., Kitzis M. D., Pialoux G., Collatz E., Gutmann L. Association of two resistance mechanisms in a clinical isolate of Enterobacter cloacae with high-level resistance to imipenem. Antimicrob Agents Chemother. 1991 Jun;35(6):1093–1098. doi: 10.1128/aac.35.6.1093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ma D., Cook D. N., Alberti M., Pon N. G., Nikaido H., Hearst J. E. Genes acrA and acrB encode a stress-induced efflux system of Escherichia coli. Mol Microbiol. 1995 Apr;16(1):45–55. doi: 10.1111/j.1365-2958.1995.tb02390.x. [DOI] [PubMed] [Google Scholar]
  25. Matsuyama S., Mizushima S. Construction and characterization of a deletion mutant lacking micF, a proposed regulatory gene for OmpF synthesis in Escherichia coli. J Bacteriol. 1985 Jun;162(3):1196–1202. doi: 10.1128/jb.162.3.1196-1202.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Misra R., Reeves P. R. Role of micF in the tolC-mediated regulation of OmpF, a major outer membrane protein of Escherichia coli K-12. J Bacteriol. 1987 Oct;169(10):4722–4730. doi: 10.1128/jb.169.10.4722-4730.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mizuno T., Chou M. Y., Inouye M. A unique mechanism regulating gene expression: translational inhibition by a complementary RNA transcript (micRNA). Proc Natl Acad Sci U S A. 1984 Apr;81(7):1966–1970. doi: 10.1073/pnas.81.7.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nakajima H., Kobayashi K., Kobayashi M., Asako H., Aono R. Overexpression of the robA gene increases organic solvent tolerance and multiple antibiotic and heavy metal ion resistance in Escherichia coli. Appl Environ Microbiol. 1995 Jun;61(6):2302–2307. doi: 10.1128/aem.61.6.2302-2307.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Okusu H., Ma D., Nikaido H. AcrAB efflux pump plays a major role in the antibiotic resistance phenotype of Escherichia coli multiple-antibiotic-resistance (Mar) mutants. J Bacteriol. 1996 Jan;178(1):306–308. doi: 10.1128/jb.178.1.306-308.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Raimondi A., Traverso A., Nikaido H. Imipenem- and meropenem-resistant mutants of Enterobacter cloacae and Proteus rettgeri lack porins. Antimicrob Agents Chemother. 1991 Jun;35(6):1174–1180. doi: 10.1128/aac.35.6.1174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Sarma V., Reeves P. Genetic locus (ompB) affecting a major outer-membrane protein in Escherichia coli K-12. J Bacteriol. 1977 Oct;132(1):23–27. doi: 10.1128/jb.132.1.23-27.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Skarstad K., Thöny B., Hwang D. S., Kornberg A. A novel binding protein of the origin of the Escherichia coli chromosome. J Biol Chem. 1993 Mar 15;268(8):5365–5370. [PubMed] [Google Scholar]
  34. Stoorvogel J., van Bussel M. J., Tommassen J., van de Klundert J. A. Molecular characterization of an Enterobacter cloacae outer membrane protein (OmpX). J Bacteriol. 1991 Jan;173(1):156–160. doi: 10.1128/jb.173.1.156-160.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wu J., Weiss B. Two divergently transcribed genes, soxR and soxS, control a superoxide response regulon of Escherichia coli. J Bacteriol. 1991 May;173(9):2864–2871. doi: 10.1128/jb.173.9.2864-2871.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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

RESOURCES