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Volume 157, Issue 5

LmrR-mediated gene regulation of multidrug resistance in Free

Abstract

Multidrug resistance (MDR) in is due to the expression of the membrane ATP-binding cassette (ABC) transporter LmrCD. In the absence of drugs, the transcriptional regulator LmrR prevents expression of the operon by binding to its operator site. Through an autoregulatory mechanism LmrR also suppresses its own expression. Although the and genes have their own promoters, primer extension analysis showed the presence of a long transcript spanning the entire – cluster, in addition to various shorter transcripts harbouring the genes only. ‘In-gel’ Cu-phenanthroline footprinting analysis indicated an extensive interaction between LmrR and the promoter/operator region. Atomic force microscopy imaging of the binding of LmrR to the control region of DNA showed severe deformations indicative of DNA wrapping and looping, while LmrR binding to a fragment containing the control region induced DNA bending. The results further suggest a drug-dependent regulation mechanism in which the genes are co-transcribed with as a polycistronic messenger. This leads to an LmrR-mediated regulation of expression that is exerted from two different locations and by distinct regulatory mechanisms.

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© 2011 SGM
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2011-05-01
2024-04-25
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References

  1. Abramoff M. D., Magelhaes P. J., Ram S. J. ( 2004). Image processing with ImageJ. Biophotonics International 11:36–42
     [Google Scholar]
  2. Agustiandari H., Lubelski J., van den Berg van Saparoea H. B., Kuipers O. P., Driessen A. J. M. ( 2008). LmrR is a transcriptional repressor of expression of the multidrug ABC transporter LmrCD in Lactococcus lactis . J Bacteriol 190:759–763 [View Article][PubMed]
     [Google Scholar]
  3. Alekshun M. N., Levy S. B. ( 1997). Regulation of chromosomally mediated multiple antibiotic resistance: the mar regulon. Antimicrob Agents Chemother 41:2067–2075[PubMed]
     [Google Scholar]
  4. Bolhuis H., Molenaar D., Poelarends G., van Veen H. W., Poolman B., Driessen A. J. M., Konings W. N. ( 1994). Proton motive force-driven and ATP-dependent drug extrusion systems in multidrug-resistant Lactococcus lactis . J Bacteriol 176:6957–6964[PubMed]
     [Google Scholar]
  5. Chopra I., Roberts M. ( 2001). Tetracycline antibiotics: mode of action, applications, molecular biology, and epidemiology of bacterial resistance. Microbiol Mol Biol Rev 65:232–260 [View Article][PubMed]
     [Google Scholar]
  6. Eckert B., Beck C. F. ( 1989). Overproduction of transposon Tn10-encoded tetracycline resistance protein results in cell death and loss of membrane potential. J Bacteriol 171:3557–3559[PubMed]
     [Google Scholar]
  7. Enoru-Eta J., Gigot D., Thia-Toong T. L., Glansdorff N., Charlier D. ( 2000). Purification and characterization of Sa-lrp, a DNA-binding protein from the extreme thermoacidophilic archaeon Sulfolobus acidocaldarius homologous to the bacterial global transcriptional regulator Lrp. J Bacteriol 182:3661–3672 [View Article][PubMed]
     [Google Scholar]
  8. Enoru-Eta J., Gigot D., Glansdorff N., Charlier D. ( 2002). High resolution contact probing of the Lrp-like DNA-binding protein Ss-Lrp from the hyperthermoacidophilic crenarchaeote Sulfolobus solfataricus P2. Mol Microbiol 45:1541–1555 [View Article][PubMed]
     [Google Scholar]
  9. Friedrich U., Lenke J. ( 2006). Improved enumeration of lactic acid bacteria in mesophilic dairy starter cultures by using multiplex quantitative real-time PCR and flow cytometry-fluorescence in situ hybridization. Appl Environ Microbiol 72:4163–4171 [View Article][PubMed]
     [Google Scholar]
  10. Gasson M. J., Kitamura Y., McLauchlan W. R., Narbad A., Parr A. J., Parsons E. L., Payne J., Rhodes M. J., Walton N. J. ( 1998). Metabolism of ferulic acid to vanillin. A bacterial gene of the enoyl-SCoA hydratase/isomerase superfamily encodes an enzyme for the hydration and cleavage of a hydroxycinnamic acid SCoA thioester. J Biol Chem 273:4163–4170 [View Article][PubMed]
     [Google Scholar]
  11. Grkovic S., Brown M. H., Skurray R. A. ( 2002). Regulation of bacterial drug export systems. Microbiol Mol Biol Rev 66:671–701 [View Article][PubMed]
     [Google Scholar]
  12. Gury J., Barthelmebs L., Tran N. P., Diviès C., Cavin J. F. ( 2004). Cloning, deletion, and characterization of PadR, the transcriptional repressor of the phenolic acid decarboxylase-encoding padA gene of Lactobacillus plantarum . Appl Environ Microbiol 70:2146–2153 [View Article][PubMed]
     [Google Scholar]
  13. Hickman R. K., McMurry L. M., Levy S. B. ( 1990). Overproduction and purification of the Tn10-specified inner membrane tetracycline resistance protein Tet using fusions to β-galactosidase. Mol Microbiol 4:1241–1251 [View Article][PubMed]
     [Google Scholar]
  14. Hochschild A., Ptashne M. ( 1986). Cooperative binding of λ repressors to sites separated by integral turns of the DNA helix. Cell 44:681–687 [View Article][PubMed]
     [Google Scholar]
  15. Hsieh P. C., Siegel S. A., Rogers B., Davis D., Lewis K. ( 1998). Bacteria lacking a multidrug pump: a sensitive tool for drug discovery. Proc Natl Acad Sci U S A 95:6602–6606 [View Article][PubMed]
     [Google Scholar]
  16. Huillet E., Velge P., Vallaeys T., Pardon P. ( 2006). LadR, a new PadR-related transcriptional regulator from Listeria monocytogenes, negatively regulates the expression of the multidrug efflux pump MdrL. FEMS Microbiol Lett 254:87–94 [View Article][PubMed]
     [Google Scholar]
  17. Lubelski J., Mazurkiewicz P., van Merkerk R., Konings W. N., Driessen A. J. M. ( 2004). ydaG and ydbA of Lactococcus lactis encode a heterodimeric ATP-binding cassette-type multidrug transporter. J Biol Chem 279:34449–34455 [View Article][PubMed]
     [Google Scholar]
  18. Lubelski J., de Jong A., van Merkerk R., Agustiandari H., Kuipers O. P., Kok J., Driessen A. J. M. ( 2006). LmrCD is a major multidrug resistance transporter in Lactococcus lactis . Mol Microbiol 61:771–781 [View Article][PubMed]
     [Google Scholar]
  19. Ma D., Alberti M., Lynch C., Nikaido H., Hearst J. E. ( 1996). The local repressor AcrR plays a modulating role in the regulation of acrAB genes of Escherichia coli by global stress signals. Mol Microbiol 19:101–112 [View Article][PubMed]
     [Google Scholar]
  20. Madoori P. K., Agustiandari H., Driessen A. J. M., Thunnissen A. M. ( 2009). Structure of the transcriptional regulator LmrR and its mechanism of multidrug recognition. EMBO J 28:156–166 [View Article][PubMed]
     [Google Scholar]
  21. Maxam A. M., Gilbert W. ( 1980). Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol 65:499–560 [View Article][PubMed]
     [Google Scholar]
  22. Mazurkiewicz P., Driessen A. J. M., Konings W. N. ( 2004). Energetics of wild-type and mutant multidrug resistance secondary transporter LmrP of Lactococcus lactis . Biochim Biophys Acta 1658:252–261 [View Article][PubMed]
     [Google Scholar]
  23. Minh P. N. L., Devroede N., Massant J., Maes D., Charlier D. ( 2009). Insights into the architecture and stoichiometry of Escherichia coli PepA*DNA complexes involved in transcriptional control and site-specific DNA recombination by atomic force microscopy. Nucleic Acids Res 37:1463–1476 [View Article][PubMed]
     [Google Scholar]
  24. Musso R. E., Di Lauro R., Adhya S., de Crombrugghe B. ( 1977). Dual control for transcription of the galactose operon by cyclic AMP and its receptor protein at two interspersed promoters. Cell 12:847–854 [View Article][PubMed]
     [Google Scholar]
  25. Neyfakh A. A., Bidnenko V. E., Chen L. B. ( 1991). Efflux-mediated multidrug resistance in Bacillus subtilis: similarities and dissimilarities with the mammalian system. Proc Natl Acad Sci U S A 88:4781–4785 [View Article][PubMed]
     [Google Scholar]
  26. Neyfakh A. A., Borsch C. M., Kaatz G. W. ( 1993). Fluoroquinolone resistance protein NorA of Staphylococcus aureus is a multidrug efflux transporter. Antimicrob Agents Chemother 37:128–129[PubMed] [CrossRef]
     [Google Scholar]
  27. Ng E. Y., Trucksis M., Hooper D. C. ( 1994). Quinolone resistance mediated by norA: physiologic characterization and relationship to flqB, a quinolone resistance locus on the Staphylococcus aureus chromosome. Antimicrob Agents Chemother 38:1345–1355[PubMed] [CrossRef]
     [Google Scholar]
  28. Overhage J., Priefert H., Steinbüchel A. ( 1999). Biochemical and genetic analyses of ferulic acid catabolism in Pseudomonas sp. strain HR199. Appl Environ Microbiol 65:4837–4847[PubMed]
     [Google Scholar]
  29. Paulsen I. T., Nguyen L., Sliwinski M. K., Rabus R., Saier M. H. Jr ( 2000). Microbial genome analyses: comparative transport capabilities in eighteen prokaryotes. J Mol Biol 301:75–100 [View Article][PubMed]
     [Google Scholar]
  30. Peeters E., Thia-Toong T. L., Gigot D., Maes D., Charlier D. ( 2004). Ss-LrpB, a novel Lrp-like regulator of Sulfolobus solfataricus P2, binds cooperatively to three conserved targets in its own control region. Mol Microbiol 54:321–336 [View Article][PubMed]
     [Google Scholar]
  31. Peeters E., Willaert R., Maes D., Charlier D. ( 2006). Ss-LrpB from Sulfolobus solfataricus condenses about 100 base pairs of its own operator DNA into globular nucleoprotein complexes. J Biol Chem 281:11721–11728 [View Article][PubMed]
     [Google Scholar]
  32. Rahmati S., Yang S., Davidson A. L., Zechiedrich E. L. ( 2002). Control of the AcrAB multidrug efflux pump by quorum-sensing regulator SdiA. Mol Microbiol 43:677–685 [View Article][PubMed]
     [Google Scholar]
  33. Rivetti C., Guthold M., Bustamante C. ( 1996). Scanning force microscopy of DNA deposited onto mica: equilibration versus kinetic trapping studied by statistical polymer chain analysis. J Mol Biol 264:919–932 [View Article][PubMed]
     [Google Scholar]
  34. Saier M. H. Jr, Paulsen I. T., Sliwinski M. K., Pao S. S., Skurray R. A., Nikaido H. ( 1998). Evolutionary origins of multidrug and drug-specific efflux pumps in bacteria. FASEB J 12:265–274[PubMed]
     [Google Scholar]
  35. Schumacher M. A., Brennan R. G. ( 2003). Deciphering the molecular basis of multidrug recognition: crystal structures of the Staphylococcus aureus multidrug binding transcription regulator QacR. Res Microbiol 154:69–77 [View Article][PubMed]
     [Google Scholar]
  36. Segura A., Bünz P. V., D’Argenio D. A., Ornston L. N. ( 1999). Genetic analysis of a chromosomal region containing vanA and vanB, genes required for conversion of either ferulate or vanillate to protocatechuate in Acinetobacter . J Bacteriol 181:3494–3504[PubMed]
     [Google Scholar]
  37. Tennent J. M., Lyon B. R., Gillespie M. T., May J. W., Skurray R. A. ( 1985). Cloning and expression of Staphylococcus aureus plasmid-mediated quaternary ammonium resistance in Escherichia coli . Antimicrob Agents Chemother 27:79–83[PubMed] [CrossRef]
     [Google Scholar]
  38. Zaidi A. H., Bakkes P. J., Lubelski J., Agustiandari H., Kuipers O. P., Driessen A. J. M. ( 2008). The ABC-type multidrug resistance transporter LmrCD is responsible for an extrusion-based mechanism of bile acid resistance in Lactococcus lactis . J Bacteriol 190:7357–7366 [View Article][PubMed]
     [Google Scholar]
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LmrR-mediated gene regulation of multidrug resistance in Lactococcus lactis
Microbiology 157, 1519 (2011); https://doi.org/10.1099/mic.0.048025-0
/content/journal/micro/10.1099/mic.0.048025-0
/content/journal/micro/10.1099/mic.0.048025-0
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