1887

Abstract

The genes of a small multidrug resistance (SMR) pump were identified as part of the pAO1-encoded nicotine regulon responsible for nicotine catabolism in . When [C]nicotine was added to the growth medium the bacteria exported the C-labelled end product of nicotine catabolism, methylamine. In the presence of the proton-motive force inhibitors 2,4-dinitrophenol (DNP), carbonyl cyanide -chlorophenylhydrazone (CCCP) or the proton ionophore nigericin, export of methylamine was inhibited and radioactivity accumulated inside the bacteria. Efflux of [C]nicotine-derived radioactivity from bacteria was also inhibited in a  :  strain with downregulated expression. Because of low amine oxidase levels in the  :  strain, --methylaminobutyrate, the methylamine precursor, accumulated. Complementation of this strain with the genes, carried on a plasmid, restored the efflux of nicotine breakdown products. Both NepA and NepB were required for full export activity, indicating that they form a two-component efflux pump. NepAB may function as a metabolic valve by exporting methylamine, the end product of nicotine catabolism, and, in conditions under which it accumulates, the intermediate --methylaminobutyrate.

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2007-05-01
2019-11-21
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References

  1. Brandsch, R. ( 2006; ). Microbiology and biochemistry of nicotine degradation. Appl Microbiol Biotechnol 69, 493–498.[CrossRef]
    [Google Scholar]
  2. Brühmüller, M. A., Schimz, A., Messmer, L. & Decker, K. ( 1975; ). Covalently bound FAD in d-6-hydroxynicotine oxidase. J Biol Chem 250, 7747–7751.
    [Google Scholar]
  3. Bugg, T., Foght, J. M., Pickard, M. A. & Gray, M. R. ( 2000; ). Uptake and active efflux of polycyclic aromatic hydrocarbons by Pseudomonas fluorescens LP6a. Appl Environ Microbiol 66, 5387–5392.[CrossRef]
    [Google Scholar]
  4. Chiribau, C.-B., Sandu, C., Fraaije, M., Schiltz, E. & Brandsch, R. ( 2004; ). A novel γ-N-methylaminobutyrate demethylating oxidase involved in catabolism of the tobacco alkaloid nicotine by Arthrobacter nicotinovorans pAO1. Eur J Biochem 271, 4677–4684.[CrossRef]
    [Google Scholar]
  5. Chiribau, C.-B., Sandu, C., Igloi, G. L. & Brandsch, R. ( 2005; ). Characterization of PmfR, the transcriptional activator of the pAO1-borne purU-mabO-folD operon of Arthrobacter nicotinovorans. J Bacteriol 187, 3062–3070.[CrossRef]
    [Google Scholar]
  6. Chiribau, C.-B., Mihasan, M., Ganas, P., Igloi, G. L., Artenie, V. & Brandsch, R. ( 2006; ). Final steps in the catabolism of nicotine. Deamination versus demethylation of γ-N-methylaminobutyrate. FEBS J 273, 1528–1536.[CrossRef]
    [Google Scholar]
  7. Daley, D. O., Rapp, M., Granseth, E., Melén, K., Drew, D. & von Heijne, G. ( 2005; ). Global topology analysis of the Escherichia coli inner membrane proteome. Science 308, 1321–1323.[CrossRef]
    [Google Scholar]
  8. Decker, K., Eberwein, H., Gries, F. A. & Brühmüller, M. ( 1961; ). Über den Abbau des Nicotins durch Bakterienenzyme. IV. L-6-Hydroxy-nicotine als erstes Zwischenprodukt. Biochem Z 334, 227–244.
    [Google Scholar]
  9. Gherna, R. L., Richardson, S. H. & Rittenberg, S. C. ( 1965; ). The bacterial oxidation of nicotine. VI. The metabolism of 2,6-dihydroxypseudooxynicotine. J Biol Chem 240, 3669–3674.
    [Google Scholar]
  10. Gottschalk, K.-E., Soskine, M., Schuldiner, S. & Kessler, H. ( 2004; ). A structural model of EmrE, a multi-drug transporter from Escherichia coli. Biophys J 86, 3335–3348.[CrossRef]
    [Google Scholar]
  11. Granseth, E., Daley, D. O., Rapp, M., Melén, K. & von Heijne, G. ( 2005; ). Experimentally constrained topology models for 51,208 bacterial inner membrane proteins. J Mol Biol 352, 489–494.[CrossRef]
    [Google Scholar]
  12. Gutman, N., Steiner-Mordoch, S. & Schuldiner, S. ( 2003; ). An amino acid cluster around the essential Glu-14 is part of the substrate- and proton-binding domain of EmrE, a multidrug transporter from Escherichia coli. J Biol Chem 278, 16082–16087.[CrossRef]
    [Google Scholar]
  13. Hearn, E. M., Dennis, J. J., Gray, M. R. & Foght, J. M. ( 2003; ). Identification and characterization of the emhABC efflux system for polycyclic aromatic hydrocarbons in Pseudomonas fluorescens cLP6a. J Bacteriol 185, 6233–6240.[CrossRef]
    [Google Scholar]
  14. Hochstein, L. I. & Rittenberg, C. S. ( 1959; ). The bacterial oxidation of nicotine. II. The isolation of the first product and its identification as (l)-6-hydroxynicotine. J Biol Chem 234, 156–162.
    [Google Scholar]
  15. Hukkanen, J., Jacob, P. & Benowitz, N. L. ( 2005; ). Metabolism and disposition kinetics of nicotine. Pharmacol Rev 57, 79–115.[CrossRef]
    [Google Scholar]
  16. Igloi, G. L. & Brandsch, R. ( 2003; ). Sequence of the 165-kilobase catabolic plasmid pAO1 from Arthrobacter nicotinovorans and identification of a pAO1-dependent nicotine uptake system. J Bacteriol 185, 1976–1989.[CrossRef]
    [Google Scholar]
  17. Jack, D. L., Storms, M. L., Tchieu, J. H., Paulsen, I. T. & Saier, M. H., Jr ( 2000; ). A broad-specificity multidrug efflux pump requiring a pair of homologous SMR-type proteins. J Bacteriol 182, 2311–2313.[CrossRef]
    [Google Scholar]
  18. Lee, G., Dallas, S., Hong, M. & Bendayan, R. ( 2001; ). Drug transport in the central nervous system: brain barriers and brain parenchyma considerations. Pharmacol Rev 53, 569–596.
    [Google Scholar]
  19. Leveau, J. H. J., Zehnder, A. J. B. & van der Meer, J. R. ( 1998; ). The tfdK gene product facilitates the uptake of 2,4-dichlorophenoxyacetate by Ralstonia eutropha JMP134 (pJP4). J Bacteriol 180, 2237–2243.
    [Google Scholar]
  20. Locher, H. H., Poolman, B., Cook, A. M. & Konings, W. N. ( 1993; ). Uptake of 4-toluene sulfonate by Comamonas testosteroni T-2. J Bacteriol 175, 1075–1080.
    [Google Scholar]
  21. Ma, C. & Chang, G. ( 2004; ). Structure of the multidrug resistance efflux transporter EmrE from Escherichia coli. Proc Natl Acad Sci U S A 101, 2852–2857.[CrossRef]
    [Google Scholar]
  22. Masaoka, Y., Ueno, Y., Morita, Y., Kuroda, T., Mizushima, T. & Tsuchiya, T. ( 2000; ). A two-component multidrug efflux pump, EbrAB, in Bacillus subtilis. J Bacteriol 182, 307–310.
    [Google Scholar]
  23. Morimyo, M., Hongo, E., Hama-Inaba, H. & Machida, I. ( 1992; ). Cloning and characterization of the mvrC gene of Escherichia coli K-12 which confers resistance against methyl viologen toxicity. Nucleic Acids Res 20, 3159–3165.[CrossRef]
    [Google Scholar]
  24. Muth, T. R. & Schuldiner, S. ( 2000; ). A membrane-embedded glutamate is required for ligand binding to the multidrug transporter EmrE. EMBO J 19, 234–240.[CrossRef]
    [Google Scholar]
  25. Nichols, N. N. & Harwood, C. S. ( 1997; ). PcaK, a high-affinity permease for the aromatic compounds 4-hydroxybenzoate and protocatechuate from Pseudomonas putida. J Bacteriol 179, 5056–5061.
    [Google Scholar]
  26. Nishino, K. & Yamaguchi, A. ( 2001; ). Analysis of a complete library of putative drug transporter genes in Escherichia coli. J Bacteriol 183, 5803–5812.[CrossRef]
    [Google Scholar]
  27. Nygaard, P. & Saxild, H. H. ( 2005; ). The purine efflux pump PbuE in Bacillus subtilis modulates expression of the PurR and G-box (XptR) regulon by adjusting the purine base pool size. J Bacteriol 187, 791–794.[CrossRef]
    [Google Scholar]
  28. Paulsen, I. T., Brown, M. H. & Skurray, R. A. ( 1996; ). Proton-dependent multidrug efflux systems. Microbiol Rev 60, 575–608.
    [Google Scholar]
  29. Pornillos, O., Chen, Y.-J., Chen, A. P. & Chang, G. ( 2005; ). X-ray structure of the EmrE multidrug transporter in complex with a substrate. Science 310, 1950–1953.[CrossRef]
    [Google Scholar]
  30. Prieto, M. A. & Garcia, J. L. ( 1997; ). Identification of the 4-hydroxyphenylacetate transporter gene of Escherichia coli W: construction of a highly sensitive cellular biosensor. FEBS Lett 414, 293–297.[CrossRef]
    [Google Scholar]
  31. Putman, M., van Veen, H. W. & Konings, W. N. ( 2000; ). Molecular properties of bacterial multidrug transporters. Microbiol Mol Biol Rev 64, 672–693.[CrossRef]
    [Google Scholar]
  32. Roduit, J.-P., Wellig, A. & Kiener, A. ( 1997; ). Renewable functionalized pyridines derived from microbial metabolites of the alkaloid (S)-nicotine. Heterocycles 45, 1687–1702.[CrossRef]
    [Google Scholar]
  33. Rojas, A., Duque, E., Mosqueda, G., Golden, G., Hurtado, A., Ramos, J. L. & Segura, A. ( 2001; ). Three efflux pumps are required to provide efficient tolerance to toluene in Pseudomonas putida DOT-T1E. J Bacteriol 183, 3967–3973.[CrossRef]
    [Google Scholar]
  34. Rotem, D., Sal-man, N. & Schuldiner, S. ( 2001; ). In vitro monomer swapping in EmrE, a multidrug transporter from Escherichia coli, reveals that the oligomer is the functional unit. J Biol Chem 276, 48243–48249.
    [Google Scholar]
  35. Sandu, C., Chiribau, C.-B. & Brandsch, R. ( 2003; ). Characterization of HdnoR, the transcriptional repressor of the 6-hydroxy-d-nicotine oxidase gene of Arthrobacter nicotinovorans pAO1, and its DNA-binding activity in response to l- and d-nicotine derivatives. J Biol Chem 278, 51307–51315.[CrossRef]
    [Google Scholar]
  36. Sandu, C., Chiribau, C.-B., Sachelaru, P. & Brandsch, R. ( 2005; ). Plasmids for nicotine-dependent and independent gene expression in Arthrobacter nicotinovorans and other Arthrobacter species. Appl Environ Microbiol 71, 8920–8924.[CrossRef]
    [Google Scholar]
  37. Schuldiner, S., Granot, D., Mordoch, S. S., Nino, S., Rotem, D., Soskin, M., Tate, C. G. & Yerushalmi, H. ( 2001; ). Small is mighty: EmrE, a multidrug transporter as an experimental paradigm. News Physiol Sci 16, 130–134.
    [Google Scholar]
  38. Ubarretxena-Belandia, I., Baldwin, J. M., Schuldiner, S. & Tate, C. G. ( 2003; ). Three-dimensional structure of the bacterial multidrug transporter EmrE shows it is an asymmetric homodimer. EMBO J 22, 6175–6181.[CrossRef]
    [Google Scholar]
  39. van Dyk, T. K., Templeton, L. J., Cantera, K. A., Sharpe, P. L. & Sariaslani, F. S. ( 2004; ). Characterization of the Escherichia coli AaeAB efflux pump: a metabolic valve? J Bacteriol 186, 7196–7204.[CrossRef]
    [Google Scholar]
  40. Yerushalmi, H., Lebendiker, M. & Schuldiner, S. ( 1995; ). EmrE, an Escherichia coli 12-kDa multidrug transporter, exchanges toxic cations and H+ and is soluble in organic solvents. J Biol Chem 270, 6856–6863.[CrossRef]
    [Google Scholar]
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