Abstract
The emrAB locus of Escherichia coli encodes a multidrug resistance pump that protects the cell from several chemically unrelated antimicrobial agents, e.g., the protonophores carbonyl cyanide m-chlorophenylhydrazone (CCCP) and tetrachlorosalicyl anilide and the antibiotics nalidixic acid and thiolactomycin. The mprA gene is located immediately upstream of this locus and was shown to be a repressor of microcin biosynthesis (I. del Castillo, J. M. Gomez, and F. Moreno, J. Bacteriol. 173:3924-3929, 1991). There is a putative transcriptional terminator sequence between the mprA and emrA genes. To locate the emr promoter, single-copy lacZ operon fusions containing different regions of the emr locus were made. Only fusions containing the mprA promoter region were expressed. mprA is thus the first gene of the operon, and we propose that it be renamed emrR. Overproduction of the EmrR protein (with a multicopy vector containing the cloned emrR gene) suppressed transcription of the emr locus. A mutation in the emrR gene led to overexpression of the EmrAB pump and increased resistance to antimicrobial agents. CCCP, nalidixic acid, and a number of other structurally unrelated chemicals induced expression of the emr genes, and the induction required EmrR. We conclude that emrRAB genes constitute an operon and that EmrR serves as a negative regulator of this operon. Some of the chemicals that induce the pump serve as its substrates, suggesting that their extrusion is the natural function of the pump.
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- Ahmed M., Borsch C. M., Taylor S. S., Vázquez-Laslop N., Neyfakh A. A. A protein that activates expression of a multidrug efflux transporter upon binding the transporter substrates. J Biol Chem. 1994 Nov 11;269(45):28506–28513. [PubMed] [Google Scholar]
- 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]
- Cohen S. P., Levy S. B., Foulds J., Rosner J. L. Salicylate induction of antibiotic resistance in Escherichia coli: activation of the mar operon and a mar-independent pathway. J Bacteriol. 1993 Dec;175(24):7856–7862. doi: 10.1128/jb.175.24.7856-7862.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dinh T., Paulsen I. T., Saier M. H., Jr A family of extracytoplasmic proteins that allow transport of large molecules across the outer membranes of gram-negative bacteria. J Bacteriol. 1994 Jul;176(13):3825–3831. doi: 10.1128/jb.176.13.3825-3831.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Furukawa H., Tsay J. T., Jackowski S., Takamura Y., Rock C. O. Thiolactomycin resistance in Escherichia coli is associated with the multidrug resistance efflux pump encoded by emrAB. J Bacteriol. 1993 Jun;175(12):3723–3729. doi: 10.1128/jb.175.12.3723-3729.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Grinius L. L., Goldberg E. B. Bacterial multidrug resistance is due to a single membrane protein which functions as a drug pump. J Biol Chem. 1994 Nov 25;269(47):29998–30004. [PubMed] [Google Scholar]
- Grinius L., Dreguniene G., Goldberg E. B., Liao C. H., Projan S. J. A staphylococcal multidrug resistance gene product is a member of a new protein family. Plasmid. 1992 Mar;27(2):119–129. doi: 10.1016/0147-619x(92)90012-y. [DOI] [PubMed] [Google Scholar]
- Jackowski S., Murphy C. M., Cronan J. E., Jr, Rock C. O. Acetoacetyl-acyl carrier protein synthase. A target for the antibiotic thiolactomycin. J Biol Chem. 1989 May 5;264(13):7624–7629. [PubMed] [Google Scholar]
- Levy S. B. Active efflux mechanisms for antimicrobial resistance. Antimicrob Agents Chemother. 1992 Apr;36(4):695–703. doi: 10.1128/aac.36.4.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lewis K. Multidrug resistance pumps in bacteria: variations on a theme. Trends Biochem Sci. 1994 Mar;19(3):119–123. doi: 10.1016/0968-0004(94)90204-6. [DOI] [PubMed] [Google Scholar]
- Libby S. J., Goebel W., Ludwig A., Buchmeier N., Bowe F., Fang F. C., Guiney D. G., Songer J. G., Heffron F. A cytolysin encoded by Salmonella is required for survival within macrophages. Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):489–493. doi: 10.1073/pnas.91.2.489. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Littlejohn T. G., Paulsen I. T., Gillespie M. T., Tennent J. M., Midgley M., Jones I. G., Purewal A. S., Skurray R. A. Substrate specificity and energetics of antiseptic and disinfectant resistance in Staphylococcus aureus. FEMS Microbiol Lett. 1992 Aug 15;74(2-3):259–265. doi: 10.1016/0378-1097(92)90439-u. [DOI] [PubMed] [Google Scholar]
- Liu J., Turnbough C. L., Jr Effects of transcriptional start site sequence and position on nucleotide-sensitive selection of alternative start sites at the pyrC promoter in Escherichia coli. J Bacteriol. 1994 May;176(10):2938–2945. doi: 10.1128/jb.176.10.2938-2945.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lomovskaya O. L., Kidwell J. P., Matin A. Characterization of the sigma 38-dependent expression of a core Escherichia coli starvation gene, pexB. J Bacteriol. 1994 Jul;176(13):3928–3935. doi: 10.1128/jb.176.13.3928-3935.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lomovskaya O., Lewis K. Emr, an Escherichia coli locus for multidrug resistance. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):8938–8942. doi: 10.1073/pnas.89.19.8938. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ma D., Cook D. N., Alberti M., Pon N. G., Nikaido H., Hearst J. E. Molecular cloning and characterization of acrA and acrE genes of Escherichia coli. J Bacteriol. 1993 Oct;175(19):6299–6313. doi: 10.1128/jb.175.19.6299-6313.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marger M. D., Saier M. H., Jr A major superfamily of transmembrane facilitators that catalyse uniport, symport and antiport. Trends Biochem Sci. 1993 Jan;18(1):13–20. doi: 10.1016/0968-0004(93)90081-w. [DOI] [PubMed] [Google Scholar]
- Marklund B. I., Tennent J. M., Garcia E., Hamers A., Båga M., Lindberg F., Gaastra W., Normark S. Horizontal gene transfer of the Escherichia coli pap and prs pili operons as a mechanism for the development of tissue-specific adhesive properties. Mol Microbiol. 1992 Aug;6(16):2225–2242. doi: 10.1111/j.1365-2958.1992.tb01399.x. [DOI] [PubMed] [Google Scholar]
- Naroditskaya V., Schlosser M. J., Fang N. Y., Lewis K. An E. coli gene emrD is involved in adaptation to low energy shock. Biochem Biophys Res Commun. 1993 Oct 29;196(2):803–809. doi: 10.1006/bbrc.1993.2320. [DOI] [PubMed] [Google Scholar]
- Neyfakh A. A., Bidnenko V. E., Chen L. B. Efflux-mediated multidrug resistance in Bacillus subtilis: similarities and dissimilarities with the mammalian system. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4781–4785. doi: 10.1073/pnas.88.11.4781. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nikaido H. Prevention of drug access to bacterial targets: permeability barriers and active efflux. Science. 1994 Apr 15;264(5157):382–388. doi: 10.1126/science.8153625. [DOI] [PubMed] [Google Scholar]
- Pabo C. O., Sauer R. T. Transcription factors: structural families and principles of DNA recognition. Annu Rev Biochem. 1992;61:1053–1095. doi: 10.1146/annurev.bi.61.070192.005201. [DOI] [PubMed] [Google Scholar]
- Paulsen I. T., Littlejohn T. G., Rådström P., Sundström L., Sköld O., Swedberg G., Skurray R. A. The 3' conserved segment of integrons contains a gene associated with multidrug resistance to antiseptics and disinfectants. Antimicrob Agents Chemother. 1993 Apr;37(4):761–768. doi: 10.1128/aac.37.4.761. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Perego M., Hoch J. A. Sequence analysis and regulation of the hpr locus, a regulatory gene for protease production and sporulation in Bacillus subtilis. J Bacteriol. 1988 Jun;170(6):2560–2567. doi: 10.1128/jb.170.6.2560-2567.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Poole K., Krebes K., McNally C., Neshat S. Multiple antibiotic resistance in Pseudomonas aeruginosa: evidence for involvement of an efflux operon. J Bacteriol. 1993 Nov;175(22):7363–7372. doi: 10.1128/jb.175.22.7363-7372.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reverchon S., Nasser W., Robert-Baudouy J. pecS: a locus controlling pectinase, cellulase and blue pigment production in Erwinia chrysanthemi. Mol Microbiol. 1994 Mar;11(6):1127–1139. doi: 10.1111/j.1365-2958.1994.tb00389.x. [DOI] [PubMed] [Google Scholar]
- Roper D. I., Fawcett T., Cooper R. A. The Escherichia coli C homoprotocatechuate degradative operon: hpc gene order, direction of transcription and control of expression. Mol Gen Genet. 1993 Feb;237(1-2):241–250. doi: 10.1007/BF00282806. [DOI] [PubMed] [Google Scholar]
- Rouch D. A., Cram D. S., DiBerardino D., Littlejohn T. G., Skurray R. A. Efflux-mediated antiseptic resistance gene qacA from Staphylococcus aureus: common ancestry with tetracycline- and sugar-transport proteins. Mol Microbiol. 1990 Dec;4(12):2051–2062. doi: 10.1111/j.1365-2958.1990.tb00565.x. [DOI] [PubMed] [Google Scholar]
- Simons R. W., Houman F., Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987;53(1):85–96. doi: 10.1016/0378-1119(87)90095-3. [DOI] [PubMed] [Google Scholar]
- Tennent J. M., Lyon B. R., Midgley M., Jones I. G., Purewal A. S., Skurray R. A. Physical and biochemical characterization of the qacA gene encoding antiseptic and disinfectant resistance in Staphylococcus aureus. J Gen Microbiol. 1989 Jan;135(1):1–10. doi: 10.1099/00221287-135-1-1. [DOI] [PubMed] [Google Scholar]
- Winans S. C., Elledge S. J., Krueger J. H., Walker G. C. Site-directed insertion and deletion mutagenesis with cloned fragments in Escherichia coli. J Bacteriol. 1985 Mar;161(3):1219–1221. doi: 10.1128/jb.161.3.1219-1221.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- del Castillo I., González-Pastor J. E., San Millán J. L., Moreno F. Nucleotide sequence of the Escherichia coli regulatory gene mprA and construction and characterization of mprA-deficient mutants. J Bacteriol. 1991 Jun;173(12):3924–3929. doi: 10.1128/jb.173.12.3924-3929.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- del Castillo I., Gómez J. M., Moreno F. mprA, an Escherichia coli gene that reduces growth-phase-dependent synthesis of microcins B17 and C7 and blocks osmoinduction of proU when cloned on a high-copy-number plasmid. J Bacteriol. 1990 Jan;172(1):437–445. doi: 10.1128/jb.172.1.437-445.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]