1887

Microbiology Society logo

Volume 147, Issue 1

Multiplicity of ammonium uptake systems in : role of Amt and AmtB Free

Abstract

In , a Gram-positive soil bacterium widely used in the industrial production of amino acids, two genes encoding (putative) ammonium uptake carriers have been described. The isolation of was the first report of the sequence of a gene encoding a bacterial ammonium uptake system combined with the characterization of the corresponding protein. Recently, a second gene, , with so far unknown function, was isolated. The isolation of this gene and the suggestion of a new concept for ammonium acquisition prompted the reinvestigation of ammonium transport in . In this study it is shown that Amt mediates uptake of (methyl)ammonium into the cell with high affinity and strictly depending on the membrane potential. As shown by the determination of at different pH values, ammonium/methylammonium, but not ammonia/methylamine, are substrates of Amt. AmtB exclusively accepts ammonium as a transport substrate. In addition, hints of another, until now unknown, low-affinity, ammonium-specific uptake system were found.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): ammonium transport , CCCP, carbonyl cyanide m-chlorophenylhydrazone. , Corynebacterium glutamicum and glutamine synthetase
© Microbiology Society
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-147-1-135
2001-01-01
2024-04-26
Loading full text...

Full text loading...

/deliver/fulltext/micro/147/1/1470135a.html?itemId=/content/journal/micro/10.1099/00221287-147-1-135&mimeType=html&fmt=ahah

References

  1. Abe, S., Takayama, K. & Kinoshita, S.(1967). Taxonomical studies on glutamic acid-producing bacteria. J Gen Microbiol 13, 279-301.[CrossRef] [Google Scholar]
  2. Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A. & Struhl, K. (1987).Current Protocols in Molecular Biology. New York: Greene Publishing Associates and Wiley Interscience.
  3. Cordts, M. L. & Gibson, J.(1987). Ammonium and methylammonium transport in Rhodobacter sphaeroides. J Bacteriol 169, 1632-1638. [Google Scholar]
  4. Cremer, J., Eggeling, L. & Sahm, H.(1990). Cloning the dapA dapB cluster of the lysine-secreting bacterium Corynebacterium glutamicum. Mol Gen Genet 220, 478-480.[CrossRef] [Google Scholar]
  5. Gazzarrini, S., Lejay, L., Gojon, A., Ninnemann, O. & Frommer, W. B.(1999). Three functional transporters for constitutive, diurnally regulated, and starvation-induced uptake of ammonium into Arabidopsis roots. Plant Cell 11, 937-947.[CrossRef] [Google Scholar]
  6. Grant, S. N. G., Jessee, J., Bloom, F. R. & Hanahan, D.(1990). Differential plasmid rescue from transgenic mouse DNAs into Escherichia coli methylation-restriction mutants. Proc Natl Acad Sci USA 87, 4645-4649.[CrossRef] [Google Scholar]
  7. Jakoby, M., Tesch, M., Sahm, H., Krämer, R. & Burkovski, A.(1997). Isolation of the Corynebacterium glutamicum glnA gene encoding glutamine synthetase I. FEMS Microbiol Lett 154, 81-88.[CrossRef] [Google Scholar]
  8. Jakoby, M., Krämer, R. & Burkovski, A.(1999). Nitrogen regulation in Corynebacterium glutamicum: isolation of genes involved and biochemical characterization of the corresponding proteins. FEMS Microbiol Lett 173, 303-310.[CrossRef] [Google Scholar]
  9. Jakoby, M., Nolden, L., Meier-Wagner, J., Krämer, R. & Burkovski, A.(2000). AmtR, a global repressor in the nitrogen regulation system of Corynebacterium glutamicum. Mol Microbiol 37, 964-977.[CrossRef] [Google Scholar]
  10. Jayakumar, A., Hong, J.-S. & Barnes, E. M.Jr(1986). Feedback inhibition of ammonium (methylammonium) ion transport in Escherichia coli by glutamine and glutamine analogs. J Bacteriol 169, 553-557. [Google Scholar]
  11. Keilhauer, C., Eggeling, L. & Sahm, H.(1993). Isoleucine synthesis in Corynebacterium glutamicum: molecular analysis of the ilvB-ilvN-ilvC operon. J Bacteriol 175, 5595-5603. [Google Scholar]
  12. Kleiner, D.(1985). Bacterial ammonium transport. FEMS Microbiol Rev 32, 87-100.[CrossRef] [Google Scholar]
  13. Kleiner, D.(1993). transport systems. In Alkali Cation Transport Systems in Prokaryotes , pp. 379-395. Edited by E. P. Bakker. Boca Raton, FL:CRC Press.
  14. Kleiner, D. & Castorph, H.(1982). Inhibition of ammonium (methylammonium) transport in Klebsiella pneumoniae by glutamine and glutamine analogues. FEBS Lett 146, 201-203.[CrossRef] [Google Scholar]
  15. Krämer, R. & Lambert, C.(1990). Uptake of glutamate in Corynebacterium glutamicum. 2. Evidence for a primary active transport system. Eur J Biochem 194, 937-944.[CrossRef] [Google Scholar]
  16. Krämer, R., Lambert, C., Hoischen, C. & Ebbighausen, H.(1990). Uptake of glutamate in Corynebacterium glutamicum. 1. Kinetic properties and regulation by internal pH and potassium. Eur J Biochem 194, 929-935.[CrossRef] [Google Scholar]
  17. Large, P. J.(1980). Microbial growth on methylated amines. In Microbial Growth on C 1 Compounds , pp. 55-69. Edited by H. Dalton. London/Philadelphia/Rheine:Heyden.
  18. Lorenz, M. C. & Heitman, J.(1998). The MEP2 ammonium permease regulates pseudohyphal differentiation in Saccharomyces cerevisiae. EMBO J 17, 1236-1247.[CrossRef] [Google Scholar]
  19. Marini, A.-M., Springael, J.-Y., Frommer, W. B. & André, B.(2000). Cross-talk between ammonium transporters in yeast and interference by the soybean SAT1 protein. Mol Microbiol 35, 378-385.[CrossRef] [Google Scholar]
  20. Montesinos, M. L., Muro-Pastor, A. M., Herrero, A. & Flores, E.(1998). Ammonium/methylammonium permeases of a cyanobacterium. J Biol Chem 273, 31463-31470.[CrossRef] [Google Scholar]
  21. Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989).Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  22. Schäfer, A., Tauch, A., Jäger, W., Kalinowski, J., Thierbach, G. & Pühler, A.(1994). Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum. Gene 145, 69-73.[CrossRef] [Google Scholar]
  23. Siewe, R. M., Weil, B., Burkovski, A., Eikmanns, B. J., Eikmanns, M. & Krämer, R.(1996). Functional and genetic characterization of the (methyl)ammonium uptake carrier of Corynebacterium glutamicum. J Biol Chem 271, 5398-5403.[CrossRef] [Google Scholar]
  24. Simon, R., Priefer, U. & Pühler, A.(1983). A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram-negative bacteria. Appl Microbiol Biotechnol 1, 784-791. [Google Scholar]
  25. Sohlenkamp, C., Shelden, M., Howitt, S. & Udvardi, M.(2000). Characterization of Arabidopsis AtAMT2, a novel ammonium transporter in plants. FEBS Lett 467, 273-278.[CrossRef] [Google Scholar]
  26. Soupene, E., He, L., Yan, D. & Kustu, S.(1998). Ammonia acquisition in enteric bacteria: physiological role of the ammonium/methylammonium transport B (AmtB) protein. Proc Natl Acad Sci USA 95, 7030-7034.[CrossRef] [Google Scholar]
  27. Tachiki, T., Wakisaka, S., Suzuki, H., Kumagai, H. & Tochikura, T.(1981). Glutamine synthetase from Micrococcus glutamicus: effect of nitrogen sources in culture medium on enzyme formation and some properties of crystalline enzyme. Agric Biol Chem 45, 287-292. [Google Scholar]
  28. Tachiki, T., Wakisaka, S., Suzuki, H., Kumagai, H. & Tochikura, T.(1983). Variation of Micrococcus glutamicus glutamine synthetase brought about by divalent cations. Agric Biol Chem 47, 287-292.[CrossRef] [Google Scholar]
  29. Tesch, M., de Graaf, A. A. & Sahm, H.(1999).In vivo fluxes in the ammonium-assimilatory pathways in Corynebacterium glutamicum studied by 15N nuclear magnetic resonance. Appl Environ Microbiol 65, 1099-1109. [Google Scholar]
  30. Thomas, G., Coutts, G. & Merrick, M.(2000). The glnKamtB operon, a conserved gene pair in prokaryotes. Trends Genet 16, 11-14. [Google Scholar]
  31. von Wirén, N., Gazzarrini, S., Gojon, A. & Frommer, W. B.(2000). The molecular physiology of ammonium uptake and retrieval. Curr Opin Plant Biol 3, 254-261.[CrossRef] [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-147-1-135
Loading
Multiplicity of ammonium uptake systems in Corynebacterium glutamicum: role of Amt and AmtB
Microbiology 147, 135 (2001); https://doi.org/10.1099/00221287-147-1-135
/content/journal/micro/10.1099/00221287-147-1-135
/content/journal/micro/10.1099/00221287-147-1-135
Loading

Data & Media loading...

Most cited Most Cited RSS feed

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