Nitrogen assimilation in Corynebacterium diphtheriae: pathways and regulatory cascades
Lars Nolden
Institute of Biochemistry, University of Cologny, Zülpicher-Str. 47, D-50674 Cologne, Germany
Search for more papers by this authorGabriele Beckers
Institute of Biochemistry, University of Cologny, Zülpicher-Str. 47, D-50674 Cologne, Germany
Search for more papers by this authorCorresponding Author
Andreas Burkovski
Institute of Biochemistry, University of Cologny, Zülpicher-Str. 47, D-50674 Cologne, Germany
*Corresponding author. Tel.: +49 (221) 470 6472; Fax: +49 (221) 470 5091, E-mail: [email protected]Search for more papers by this authorLars Nolden
Institute of Biochemistry, University of Cologny, Zülpicher-Str. 47, D-50674 Cologne, Germany
Search for more papers by this authorGabriele Beckers
Institute of Biochemistry, University of Cologny, Zülpicher-Str. 47, D-50674 Cologne, Germany
Search for more papers by this authorCorresponding Author
Andreas Burkovski
Institute of Biochemistry, University of Cologny, Zülpicher-Str. 47, D-50674 Cologne, Germany
*Corresponding author. Tel.: +49 (221) 470 6472; Fax: +49 (221) 470 5091, E-mail: [email protected]Search for more papers by this authorAbstract
Genes encoding proteins for ammonium uptake, assimilation, and the nitrogen regulatory system in Corynebacterium diphtheriae were studied on basis of homology searches using Corynebacterium glutamicum genes as query sequences. Regulation of transcription of these genes in response to nitrogen starvation was analyzed by RNA hybridization experiments and knock-out mutants were generated to verify the function of distinct genes. In this communication, we were able to identify the key components of ammonium assimilation pathways and nitrogen regulation in C. diphtheriae. Moreover, we show in this study that molecular biology methods and vectors developed for C. glutamicum can be applied in C. diphtheriae. The results obtained strengthens the role of C. glutamicum as a model organism for mycolic acids-containing actinomycetes.
References
- [1] Beckers, G., Nolden, L., Burkovski, A. (2001) Glutamate synthase of Corynebacterium glutamicum is not essential for glutamate synthesis and is regulated by the nitrogen status. Microbiology 147, 2961–2970.
- [2] Clemens, D.L., Lee, B.-Y., Horwitz, M.A. (1995) Purification, characterization, and genetic analysis of Mycobacterium tuberculosis urease, a potentially critical determinant of host–pathogen interaction. J. Bacteriol 177, 5644–5652.
- [3] Eaton, K.A., Brooks, C.L., Morgan, D.R., Krakowka, S. (1991) Essential role of urease in pathogenesis of gastritis induced by Helicobacter pylori in gnotobiotic piglets. Infect. Immun 59, 2470–2475.
- [4] Eikmanns, B.J., Thum-Schmitz, N., Eggeling, L., Lüdtke, K.-U., Sahm, H. (1994) Nucleotide sequence, expression and transcriptional analysis of the Corynebacterium glutamicum gltA gene encoding citrate synthase. Microbiology 140, 1817–1828.
- [5] Funke, G., von Graevenitz, A. J.E. Clarridge III Bernard, K. (1997) Clinical microbiology of coryneform bacteria. Clin. Microbiol. Rev 10, 125–159.
- [6] Funke, G., Altwegg, M., Frommelt, L., von Graevenitz, A. (1999) Emergence of related non-toxogenic Corynebacterium diphtheriae biotype mitis strains in western Europe. Emerg. Infect. Dis 5, 477–480.
- [7] Harth, G., Horwitz, M.A. (1999) An inhibitor of exported Mycobacterium tuberculosis: glutamine synthetase selectively blocks growth of pathogenic mycobacteria in axenic culture and in human monocytes: extracellular proteins as a potential drug target. J. Exp. Med 189, 1425–1435.
- [8] Harth, G., Clemens, D.L., Horwitz, M.A. (1994) Glutamine synthetase of Mycobacterium tuberculosis: extracellular release and characterization of its enzymatic activity. Proc. Natl. Acad. Sci. USA 91, 9342–9346.
- [9] Hogg, G.G., Strachan, J.E., Huayi, L., Beaton, S.A., Robinson, P.M., Taylor, K. (1996) Non-toxigenic Corynebacterium diphtheriae biovar gravis: evidence for an invasive clone in south-eastern Australian community. Med. J. Aust 164, 72–75.
- [10] Jakoby, M., Krämer, R., Burkovski, A. (1999) Nitrogen regulation in Corynebacterium glutamicum: isolation of genes involved and biochemical characterization of corresponding proteins. FEMS Microbiol. Lett 173, 303–310.
- [11] 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.
- [12] Liebl, W., Bayerl, A., Schein, B., Stillner, U., Schleifer, K.H. (1989) High efficiency electroporation of intact Corynebacterium glutamicum cells. FEMS Microbiol. Lett 53, 299–303.
- [13] Martinez-Martinez, L. (1998) Clinical significance of newly recognized coryneform bacteria. Rev. Med. Microbiol 9, 55–68.
- [14] Meier-Wagner, J., Nolden, L., Jakoby, M., Siewe, R., Krämer, R., Burkovski, A. (2001) Multiplicity of ammonium uptake systems in Corynebacterium glutamicum: role of Amt and AmtB. Microbiology 147, 135–143.
- [15] Nolden, L., Farwick, M., Krämer, R., Burkovski, A. (2001) Glutamine synthetases in Corynebacterium glutamicum: transcriptional control and regulation of activity. FEMS Microbiol. Lett 201, 91–98.
- [16] Nolden, L., Ngouoto-Nkili, C.-E., Bendt, A.K., Krämer, R., Burkovski, A. (2001) Sensing nitrogen limitation in Corynebacterium glutamicum: the role of glnK and glnD. Mol. Microbiol 42, 1281–1295.
- [17] Sambrook, J., Fritsch, E.F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd edition. Cold Spring Habor Laboratory Press, Cold Spring Habor, NY.
- [18] 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.
- [19] Schmitt, M.P., Drazek, E.S. (2001) Construction and consequences of directed mutations affecting the hemin receptor in pathogenic Corynebacterium species. J. Bacteriol 183, 1476–1481.
- [20] 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.
- [21] Thomas, G., Coutts, G., Merrick, M. (2000) The glnKamtB operon, a conserved gene pair in prokaryotes. Trends Genet 16, 11–14.
- [22] Vitek, C.R., Wharton, M. (1998) Diphteriae in the former Soviet Union: reemergence of pandemic disease. Emerg. Infect. Dis 4, 539–550.
- [23] Wakisaka, S., Tachiki, T., Tochikura, T. (1990) Properties of Brevibacterium flavum glutamine synthetase in an ‘in-vivo-like’ system. J. Ferm. Bioeng 70, 182–184.
- [24] Yanisch-Perron, C., Vieira, L., Messing, J. (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of M13mp18 and pUC19 vectors. Gene 33, 103–119.
