Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1993 Feb;175(4):1026–1031. doi: 10.1128/jb.175.4.1026-1031.1993

Posttranscriptional repression of Escherichia coli OmpF protein in response to redox stress: positive control of the micF antisense RNA by the soxRS locus.

J H Chou 1, J T Greenberg 1, B Demple 1
PMCID: PMC193015  PMID: 7679383

Abstract

The soxRS regulon is a cornerstone of the adaptive defense systems of Escherichia coli against oxidative stress. Unexpectedly, activation of this regulon also enhances bacterial resistance to multiple antibiotics that seem unrelated to oxygen radicals. We previously correlated this multiple antibiotic resistance with a reduced rate of synthesis of the OmpF outer membrane porin that does not affect the OmpC or OmpA porins. Studies presented here, with operon and gene fusions of ompF to lacZ, show that the soxRS-dependent repression of OmpF is achieved posttranscriptionally. We also show posttranscriptional repression of OmpF mediated by the soxQ1 mutation, which maps to the marA locus. These repressions are dependent on the micF gene, which encodes a small RNA partially complementary to the 5' end of the ompF message. Northern (RNA) blotting experiments show that micF transcription is strongly inducible by the superoxide-generating agent paraquat in a manner that depends completely on the soxRS locus. The soxR-constitutive and soxQ1 mutations elevate the expression of micF in the absence of redox stress. However, the antibiotic resistance mediated by a soxR-constitutive mutation was only partially reversed upon deletion of micF. The soxRS regulon therefore includes other components that contribute to general antibiotic resistance, although the relation of this phenotype to oxidative stress remains to be established.

Full text

PDF
1026

Images in this article

1029Image
on p.1029

Selected References

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

  1. Amábile-Cuevas C. F., Demple B. Molecular characterization of the soxRS genes of Escherichia coli: two genes control a superoxide stress regulon. Nucleic Acids Res. 1991 Aug 25;19(16):4479–4484. doi: 10.1093/nar/19.16.4479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andersen J., Delihas N. micF RNA binds to the 5' end of ompF mRNA and to a protein from Escherichia coli. Biochemistry. 1990 Oct 2;29(39):9249–9256. doi: 10.1021/bi00491a020. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Carlioz A., Touati D. Isolation of superoxide dismutase mutants in Escherichia coli: is superoxide dismutase necessary for aerobic life? EMBO J. 1986 Mar;5(3):623–630. doi: 10.1002/j.1460-2075.1986.tb04256.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen S. P., Hooper D. C., Wolfson J. S., Souza K. S., McMurry L. M., Levy S. B. Endogenous active efflux of norfloxacin in susceptible Escherichia coli. Antimicrob Agents Chemother. 1988 Aug;32(8):1187–1191. doi: 10.1128/aac.32.8.1187. [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. Csonka L. N. Physiological and genetic responses of bacteria to osmotic stress. Microbiol Rev. 1989 Mar;53(1):121–147. doi: 10.1128/mr.53.1.121-147.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cunningham R. P., Saporito S. M., Spitzer S. G., Weiss B. Endonuclease IV (nfo) mutant of Escherichia coli. J Bacteriol. 1986 Dec;168(3):1120–1127. doi: 10.1128/jb.168.3.1120-1127.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Demple B. Regulation of bacterial oxidative stress genes. Annu Rev Genet. 1991;25:315–337. doi: 10.1146/annurev.ge.25.120191.001531. [DOI] [PubMed] [Google Scholar]
  11. Garvey N., St John A. C., Witkin E. M. Evidence for RecA protein association with the cell membrane and for changes in the levels of major outer membrane proteins in SOS-induced Escherichia coli cells. J Bacteriol. 1985 Sep;163(3):870–876. doi: 10.1128/jb.163.3.870-876.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Genilloud O., Garrido M. C., Moreno F. The transposon Tn5 carries a bleomycin-resistance determinant. Gene. 1984 Dec;32(1-2):225–233. doi: 10.1016/0378-1119(84)90050-7. [DOI] [PubMed] [Google Scholar]
  13. George A. M., Levy S. B. Amplifiable resistance to tetracycline, chloramphenicol, and other antibiotics in Escherichia coli: involvement of a non-plasmid-determined efflux of tetracycline. J Bacteriol. 1983 Aug;155(2):531–540. doi: 10.1128/jb.155.2.531-540.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. George A. M., Levy S. B. Gene in the major cotransduction gap of the Escherichia coli K-12 linkage map required for the expression of chromosomal resistance to tetracycline and other antibiotics. J Bacteriol. 1983 Aug;155(2):541–548. doi: 10.1128/jb.155.2.541-548.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Greenberg J. T., Chou J. H., Monach P. A., Demple B. Activation of oxidative stress genes by mutations at the soxQ/cfxB/marA locus of Escherichia coli. J Bacteriol. 1991 Jul;173(14):4433–4439. doi: 10.1128/jb.173.14.4433-4439.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Greenberg J. T., Demple B. A global response induced in Escherichia coli by redox-cycling agents overlaps with that induced by peroxide stress. J Bacteriol. 1989 Jul;171(7):3933–3939. doi: 10.1128/jb.171.7.3933-3939.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Greenberg J. T., Monach P., Chou J. H., Josephy P. D., Demple B. Positive control of a global antioxidant defense regulon activated by superoxide-generating agents in Escherichia coli. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6181–6185. doi: 10.1073/pnas.87.16.6181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. Hitz H., Schafer D., Wittmann-Liebold B. Primary structure of ribosomal protein S6 from the wild type and a mutant of Escherichia coli. FEBS Lett. 1975 Aug 15;56(2):259–262. doi: 10.1016/0014-5793(75)81105-7. [DOI] [PubMed] [Google Scholar]
  21. Hooper D. C., Wolfson J. S., Bozza M. A., Ng E. Y. Genetics and regulation of outer membrane protein expression by quinolone resistance loci nfxB, nfxC, and cfxB. Antimicrob Agents Chemother. 1992 May;36(5):1151–1154. doi: 10.1128/aac.36.5.1151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hooper D. C., Wolfson J. S., Ng E. Y., Swartz M. N. Mechanisms of action of and resistance to ciprofloxacin. Am J Med. 1987 Apr 27;82(4A):12–20. [PubMed] [Google Scholar]
  23. Hooper D. C., Wolfson J. S., Souza K. S., Ng E. Y., McHugh G. L., Swartz M. N. Mechanisms of quinolone resistance in Escherichia coli: characterization of nfxB and cfxB, two mutant resistance loci decreasing norfloxacin accumulation. Antimicrob Agents Chemother. 1989 Mar;33(3):283–290. doi: 10.1128/aac.33.3.283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hächler H., Cohen S. P., Levy S. B. marA, a regulated locus which controls expression of chromosomal multiple antibiotic resistance in Escherichia coli. J Bacteriol. 1991 Sep;173(17):5532–5538. doi: 10.1128/jb.173.17.5532-5538.1991. [DOI] [PMC free article] [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. Nunoshiba T., Hidalgo E., Amábile Cuevas C. F., Demple B. Two-stage control of an oxidative stress regulon: the Escherichia coli SoxR protein triggers redox-inducible expression of the soxS regulatory gene. J Bacteriol. 1992 Oct;174(19):6054–6060. doi: 10.1128/jb.174.19.6054-6060.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Quinlan G. J., Gutteridge J. M. Hydroxyl radical generation by the tetracycline antibiotics with free radical damage to DNA, lipids and carbohydrate in the presence of iron and copper salts. Free Radic Biol Med. 1988;5(5-6):341–348. doi: 10.1016/0891-5849(88)90106-2. [DOI] [PubMed] [Google Scholar]
  30. Quinlan G. J., Gutteridge J. M. Oxidative damage to DNA and deoxyribose by beta-lactam antibiotics in the presence of iron and copper salts. Free Radic Res Commun. 1988;5(3):149–158. doi: 10.3109/10715768809066924. [DOI] [PubMed] [Google Scholar]
  31. Rao R. N., Rogers S. G. Plasmid pKC7: a vector containing ten restriction endonuclease sites suitable for cloning DNA segments. Gene. 1979 Sep;7(1):79–82. doi: 10.1016/0378-1119(79)90044-1. [DOI] [PubMed] [Google Scholar]
  32. Rosner J. L., Chai T. J., Foulds J. Regulation of ompF porin expression by salicylate in Escherichia coli. J Bacteriol. 1991 Sep;173(18):5631–5638. doi: 10.1128/jb.173.18.5631-5638.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Skolimowski I. M., Knight R. C., Edwards D. I. Molecular basis of chloramphenicol and thiamphenicol toxicity to DNA in vitro. J Antimicrob Chemother. 1983 Dec;12(6):535–542. doi: 10.1093/jac/12.6.535. [DOI] [PubMed] [Google Scholar]
  34. Tsaneva I. R., Weiss B. soxR, a locus governing a superoxide response regulon in Escherichia coli K-12. J Bacteriol. 1990 Aug;172(8):4197–4205. doi: 10.1128/jb.172.8.4197-4205.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Vermeersch G., Ronfard-Haret J. C., Bazin M., Carillet V., Morliere P., Santus R. Type I and type II photosensitization by the antibacterial drug nalidixic acid. A laser flash photolysis study. Photochem Photobiol. 1991 Nov;54(5):661–666. doi: 10.1111/j.1751-1097.1991.tb02072.x. [DOI] [PubMed] [Google Scholar]
  36. Walkup L. K., Kogoma T. Escherichia coli proteins inducible by oxidative stress mediated by the superoxide radical. J Bacteriol. 1989 Mar;171(3):1476–1484. doi: 10.1128/jb.171.3.1476-1484.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wu J., Weiss B. Two-stage induction of the soxRS (superoxide response) regulon of Escherichia coli. J Bacteriol. 1992 Jun;174(12):3915–3920. doi: 10.1128/jb.174.12.3915-3920.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Yasuda S., Takagi T. Overproduction of Escherichia coli replication proteins by the use of runaway-replication plasmids. J Bacteriol. 1983 Jun;154(3):1153–1161. doi: 10.1128/jb.154.3.1153-1161.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES