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.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.2006/004234-0
2007-05-01
2021-07-30
Loading full text...

Full text loading...

/deliver/fulltext/micro/153/5/1546.html?itemId=/content/journal/micro/10.1099/mic.0.2006/004234-0&mimeType=html&fmt=ahah

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., Drew D., Melén K., 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., Konings W. N., van Veen H. W. 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]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.2006/004234-0
Loading
/content/journal/micro/10.1099/mic.0.2006/004234-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error