Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Oct;179(19):6122–6126. doi: 10.1128/jb.179.19.6122-6126.1997

Role of the acrAB locus in organic solvent tolerance mediated by expression of marA, soxS, or robA in Escherichia coli.

D G White 1, J D Goldman 1, B Demple 1, S B Levy 1
PMCID: PMC179517  PMID: 9324261

Abstract

Escherichia coli K-12 strains are normally tolerant to n-hexane and susceptible to cyclohexane. Constitutive expression of marA of the multiple antibiotic resistance (mar) locus or of the soxS or robA gene product produced tolerance to cyclohexane. Inactivation of the mar locus or the robA locus, but not the soxRS locus, increased organic solvent susceptibility in the wild type and Mar mutants (to both n-hexane and cyclohexane). The organic solvent hypersusceptibility is a newly described phenotype for a robA-inactivated strain. Multicopy expression of mar, soxS, or robA induced cyclohexane tolerance in strains with a deleted or inactivated chromosomal mar, soxRS, or robA locus; thus, each transcriptional activator acts independently of the others. However, in a strain with 39 kb of chromosomal DNA, including the mar locus, deleted, only the multicopy complete mar locus, consisting of its two operons, produced cyclohexane tolerance. Deletion of acrAB from either wild-type E. coli K-12 or a Mar mutant resulted in loss of tolerance to both n-hexane and cyclohexane. Organic solvent tolerance mediated by mar, soxS, or robA was not restored in strains with acrAB deleted. These findings strongly suggest that active efflux specified by the acrAB locus is linked to intrinsic organic solvent tolerance and to tolerance mediated by the marA, soxS, or robA gene product in E. coli.

Full Text

The Full Text of this article is available as a PDF (143.1 KB).

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. Aono R., Kobayashi M., Nakajima H., Kobayashi H. A close correlation between improvement of organic solvent tolerance levels and alteration of resistance toward low levels of multiple antibiotics in Escherichia coli. Biosci Biotechnol Biochem. 1995 Feb;59(2):213–218. doi: 10.1271/bbb.59.213. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. Asako H., Nakajima H., Kobayashi K., Kobayashi M., Aono R. Organic solvent tolerance and antibiotic resistance increased by overexpression of marA in Escherichia coli. Appl Environ Microbiol. 1997 Apr;63(4):1428–1433. doi: 10.1128/aem.63.4.1428-1433.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Cohen S. P., McMurry L. M., Hooper D. C., Wolfson J. S., Levy S. B. Cross-resistance to fluoroquinolones in multiple-antibiotic-resistant (Mar) Escherichia coli selected by tetracycline or chloramphenicol: decreased drug accumulation associated with membrane changes in addition to OmpF reduction. Antimicrob Agents Chemother. 1989 Aug;33(8):1318–1325. doi: 10.1128/aac.33.8.1318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Fawcett W. P., Wolf R. E., Jr Purification of a MalE-SoxS fusion protein and identification of the control sites of Escherichia coli superoxide-inducible genes. Mol Microbiol. 1994 Nov;14(4):669–679. doi: 10.1111/j.1365-2958.1994.tb01305.x. [DOI] [PubMed] [Google Scholar]
  10. Ferrante A. A., Augliera J., Lewis K., Klibanov A. M. Cloning of an organic solvent-resistance gene in Escherichia coli: the unexpected role of alkylhydroperoxide reductase. Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7617–7621. doi: 10.1073/pnas.92.17.7617. [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., 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]
  13. Goldman J. D., White D. G., Levy S. B. Multiple antibiotic resistance (mar) locus protects Escherichia coli from rapid cell killing by fluoroquinolones. Antimicrob Agents Chemother. 1996 May;40(5):1266–1269. doi: 10.1128/aac.40.5.1266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. Isken S., de Bont J. A. Active efflux of toluene in a solvent-resistant bacterium. J Bacteriol. 1996 Oct;178(20):6056–6058. doi: 10.1128/jb.178.20.6056-6058.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jackson R. W., DeMoss J. A. Effects of toluene on Escherichia coli. J Bacteriol. 1965 Nov;90(5):1420–1425. doi: 10.1128/jb.90.5.1420-1425.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jair K. W., Yu X., Skarstad K., Thöny B., Fujita N., Ishihama A., Wolf R. E., Jr Transcriptional activation of promoters of the superoxide and multiple antibiotic resistance regulons by Rob, a binding protein of the Escherichia coli origin of chromosomal replication. J Bacteriol. 1996 May;178(9):2507–2513. doi: 10.1128/jb.178.9.2507-2513.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Li Z., Demple B. SoxS, an activator of superoxide stress genes in Escherichia coli. Purification and interaction with DNA. J Biol Chem. 1994 Jul 15;269(28):18371–18377. [PubMed] [Google Scholar]
  20. Ma D., Alberti M., Lynch C., Nikaido H., Hearst J. E. The local repressor AcrR plays a modulating role in the regulation of acrAB genes of Escherichia coli by global stress signals. Mol Microbiol. 1996 Jan;19(1):101–112. doi: 10.1046/j.1365-2958.1996.357881.x. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Maneewannakul K., Levy S. B. Identification for mar mutants among quinolone-resistant clinical isolates of Escherichia coli. Antimicrob Agents Chemother. 1996 Jul;40(7):1695–1698. doi: 10.1128/aac.40.7.1695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Martin R. G., Rosner J. L. Binding of purified multiple antibiotic-resistance repressor protein (MarR) to mar operator sequences. Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5456–5460. doi: 10.1073/pnas.92.12.5456. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McMurry L. M., George A. M., Levy S. B. Active efflux of chloramphenicol in susceptible Escherichia coli strains and in multiple-antibiotic-resistant (Mar) mutants. Antimicrob Agents Chemother. 1994 Mar;38(3):542–546. doi: 10.1128/aac.38.3.542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Miller P. F., Gambino L. F., Sulavik M. C., Gracheck S. J. Genetic relationship between soxRS and mar loci in promoting multiple antibiotic resistance in Escherichia coli. Antimicrob Agents Chemother. 1994 Aug;38(8):1773–1779. doi: 10.1128/aac.38.8.1773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Nakajima H., Kobayashi M., Negishi T., Aono R. soxRS gene increased the level of organic solvent tolerance in Escherichia coli. Biosci Biotechnol Biochem. 1995 Jul;59(7):1323–1325. doi: 10.1271/bbb.59.1323. [DOI] [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. 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. Rosner J. L., Slonczewski J. L. Dual regulation of inaA by the multiple antibiotic resistance (mar) and superoxide (soxRS) stress response systems of Escherichia coli. J Bacteriol. 1994 Oct;176(20):6262–6269. doi: 10.1128/jb.176.20.6262-6269.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Seoane A. S., Levy S. B. Characterization of MarR, the repressor of the multiple antibiotic resistance (mar) operon in Escherichia coli. J Bacteriol. 1995 Jun;177(12):3414–3419. doi: 10.1128/jb.177.12.3414-3419.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Woldringh C. L. Effects of toluene and phenethyl alcohol on the ultrastructure of Escherichia coli. J Bacteriol. 1973 Jun;114(3):1359–1361. doi: 10.1128/jb.114.3.1359-1361.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. de Smet M. J., Kingma J., Witholt B. The effect of toluene on the structure and permeability of the outer and cytoplasmic membranes of Escherichia coli. Biochim Biophys Acta. 1978 Jan 4;506(1):64–80. doi: 10.1016/0005-2736(78)90435-2. [DOI] [PubMed] [Google Scholar]

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

RESOURCES