1887

Abstract

Glutamate racemase (MurI) provides -glutamate, a key building block in the peptidoglycan of the bacterial cell wall. Besides having a crucial role in cell wall biosynthesis, MurI proteins from some bacteria have been shown to act as an inhibitor of DNA gyrase. and MurI exhibit these dual characteristics. Here, we show that the two activities of MurI are unlinked and independent of each other. The racemization function of MurI is not essential for its gyrase-inhibitory property. MurI–DNA gyrase interaction influences gyrase activity but has no effect on the racemization activity of MurI. Overexpression of MurI provides resistance to the action of ciprofloxacin, suggesting the importance of the interaction in gyrase modulation. We propose that the moonlighting activity of MurI has evolved more recently than its racemase function, to play a transient yet important role in gyrase modulation.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.2008/020933-0
2008-09-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/154/9/2796.html?itemId=/content/journal/micro/10.1099/mic.0.2008/020933-0&mimeType=html&fmt=ahah

References

  1. Ashiuchi M., Kuwana E., Yamamoto T., Komatsu K., Soda K., Misono H. 2002; Glutamate racemase is an endogenous DNA gyrase inhibitor. J Biol Chem 277:39070–39073
    [Google Scholar]
  2. Ashiuchi M., Kuwana E., Komatsu K., Soda K., Misono H. 2003; Differences in effects on DNA gyrase activity between two glutamate racemases of Bacillus subtilis , the poly-gamma-glutamate synthesis-linking Glr enzyme and the YrpC (MurI) isozyme. FEMS Microbiol Lett 223:221–225
    [Google Scholar]
  3. Faik P., Walker J. I., Redmill A. A., Morgan M. J. 1988; Mouse glucose-6-phosphate isomerase and neuroleukin have identical 3′ sequences. Nature 332:455–457
    [Google Scholar]
  4. Gallo K. A., Knowles J. R. 1993; Purification, cloning, and cofactor independence of glutamate racemase from Lactobacillus . Biochemistry 32:3981–3990
    [Google Scholar]
  5. Gallo K. A., Tanner M. E., Knowles J. R. 1993; Mechanism of the reaction catalyzed by glutamate racemase. Biochemistry 32:3991–3997
    [Google Scholar]
  6. Glavas S., Tanner M. E. 1999; Catalytic acid/base residues of glutamate racemase. Biochemistry 38:4106–4113
    [Google Scholar]
  7. Glavas S., Tanner M. E. 2001; Active site residues of glutamate racemase. Biochemistry 40:6199–6204
    [Google Scholar]
  8. Henderson B. 2005 Moonlighting in Protein Hyperspace: Shared Moonlighting Proteins and Bacteria–Host Crosstalk Cambridge, UK: Cambridge University Press;
    [Google Scholar]
  9. Jeffery C. J. 1999; Moonlighting proteins. Trends Biochem Sci 24:8–11
    [Google Scholar]
  10. Jeffery C. J. 2003; Moonlighting proteins: old proteins learning new tricks. Trends Genet 19:415–417
    [Google Scholar]
  11. Kenez J. 1973; One gene – one enzyme. G. W. Beadle 70 years old. Orv Hetil 114:3031–3033 in Hungarian
    [Google Scholar]
  12. Kirsch R. D., Joly E. 1998; An improved PCR-mutagenesis strategy for two-site mutagenesis or sequence swapping between related genes. Nucleic Acids Res 26:1848–1850
    [Google Scholar]
  13. Lee Y. H., Nadaraia S., Gu D., Becker D. F., Tanner J. J. 2003; Structure of the proline dehydrogenase domain of the multifunctional PutA flavoprotein. Nat Struct Biol 10:109–114
    [Google Scholar]
  14. Lim M. L., Lum M. G., Hansen T. M., Roucou X., Nagley P. 2002; On the release of cytochrome c from mitochondria during cell death signaling. J Biomed Sci 9:488–506
    [Google Scholar]
  15. Lipinska B., Zylicz M., Georgopoulos C. 1990; The HtrA (DegP) protein, essential for Escherichia coli survival at high temperatures, is an endopeptidase. J Bacteriol 172:1791–1797
    [Google Scholar]
  16. Lundqvist T., Fisher S. L., Kern G., Folmer R. H., Xue Y., Newton D. T., Keating T. A., Alm R. A., de Jonge B. L. 2007; Exploitation of structural and regulatory diversity in glutamate racemases. Nature 447:817–822
    [Google Scholar]
  17. Manjunatha U. H., Dalal M., Chatterji M., Radha D. R., Visweswariah S. S., Nagaraja V. 2002; Functional characterisation of mycobacterial DNA gyrase: an efficient decatenase. Nucleic Acids Res 30:2144–2153
    [Google Scholar]
  18. Maxwell A., Howells A. J. 1999; Overexpression and purification of bacterial DNA gyrase. Methods Mol Biol 94:135–144
    [Google Scholar]
  19. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  20. Sengupta S., Shah M., Nagaraja V. 2006; Glutamate racemase from Mycobacterium tuberculosis inhibits DNA gyrase by affecting its DNA-binding. Nucleic Acids Res 34:5567–5576
    [Google Scholar]
  21. Spiess C., Beil A., Ehrmann M. 1999; A temperature-dependent switch from chaperone to protease in a widely conserved heat shock protein. Cell 97:339–347
    [Google Scholar]
  22. Suzuki S., Kobayashi M., Chiba K., Horiuchi I., Wang J., Kondoh T., Hashino S., Tanaka J., Hosokawa M., Asaka M. 2002; Autocrine production of epithelial cell-derived neutrophil attractant-78 induced by granulocyte colony-stimulating factor in neutrophils. Blood 99:1863–1865
    [Google Scholar]
  23. Tanaka N., Haga A., Uemura H., Akiyama H., Funasaka T., Nagase H., Raz A., Nakamura K. T. 2002; Inhibition mechanism of cytokine activity of human autocrine motility factor examined by crystal structure analyses and site-directed mutagenesis studies. J Mol Biol 318:985–997
    [Google Scholar]
  24. Triccas J. A., Parish T., Britton W. J., Gicquel B. 1998; An inducible expression system permitting the efficient purification of a recombinant antigen from Mycobacterium smegmatis . FEMS Microbiol Lett 167:151–156
    [Google Scholar]
  25. Unniraman S., Nagaraja V. 1999; Regulation of DNA gyrase operon in Mycobacterium smegmatis : a distinct mechanism of relaxation stimulated transcription. Genes Cells 4:697–706
    [Google Scholar]
  26. Watanabe H., Takehana K., Date M., Shinozaki T., Raz A. 1996; Tumor cell autocrine motility factor is the neuroleukin/phosphohexose isomerase polypeptide. Cancer Res 56:2960–2963
    [Google Scholar]
  27. Xu W., Seiter K., Feldman E., Ahmed T., Chiao J. W. 1996; The differentiation and maturation mediator for human myeloid leukemia cells shares homology with neuroleukin or phosphoglucose isomerase. Blood 87:4502–4506
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.2008/020933-0
Loading
/content/journal/micro/10.1099/mic.0.2008/020933-0
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