Skip to main content
Log in

Identification of two glutaminases inRhizobium etli

  • Published:
Biochemical Genetics Aims and scope Submit manuscript

Abstract

We present evidence thatRhizobium etli has two glutaminases differentiated by their thermostability and electrophoretic mobility. The thermostable glutaminase (B) is constitutive, in contrast with the thermolabile glutaminase (A), which is positively regulated by glutamine and negatively regulated by ammonium and by the carbon source. In distinction to glutaminase A, glutaminase B plays a minor role in the utilization of glutamine as a carbon source, but it may play a role in maintaining the balance of glutamine and glutamate. By complementation of theRhizobium etli LM16 mutant that lacks glutaminase A, we have cloned the gene that codes for this enzyme.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Aledo, J. C., Gómez-Biedma, S., Segura, J. A., Molina, M., Nuñez de Castro, I., and Márquez, J. (1993). Native gel electrophoresis of membrane proteins: Glutaminase detectionin situ specific activity staining.Electrophoresis 1488.

    Google Scholar 

  • Beringer, J. E. (1974). R factor transfer inRhizobium leguminosarum.J. Gen. Microbiol. 84198.

    Google Scholar 

  • Boyer, H. B., and Roulland-Dussoix, D. (1969). A complementation analysis of the restriction and modification of DNA inEscherichia coli.J. Mol. Biol. 41459.

    Google Scholar 

  • Calderón, J., and Martínez, L. M. (1993). Regulation of ammonium ion assimilation enzymes inNeurospora crassa nit-2 andms-5 mutant strains.Biochem. Genet. 31452.

    Google Scholar 

  • Calderón, J., and Mora, J. (1985). Glutamine cycling inNeurospora crassa.J. Gen. Microbiol. 1313237.

    Google Scholar 

  • Calderón, J., and Mora, J. (1989). Glutamine assimilation pathways inNeurospora crassa growing on glutamine as sole nitrogen and carbon source.J. Gen. Microbiol. 1352699.

    Google Scholar 

  • Calderón, J., Morett, E., and Mora, J. (1985). θ-Amidase pathway in the degradation of glutamine inNeurospora crassa.J. Bacteriol. 161807.

    Google Scholar 

  • Calderón, J., Cooper, A. J. L., Gelbard, A. S., and Mora, J. (1989).13N isotope studies of glutamine assimilation pathways inNeurospora crassa.J. Bacteriol. 1711772.

    Google Scholar 

  • Chaney, A. L., and Marbach, E. P. (1962). Modified reagents for the determination of urea and ammonia.Clin. Chem. 8130.

    Google Scholar 

  • Cooper, A. J. L., and Meister, A. (1977). The glutamine transaminase-θ-amidase pathway.Crit. Rev. Biochem. 4281.

    Google Scholar 

  • Corbin, D., Ditta, G., and Helinski, R. (1982). Clustering of nitrogen fixation (nif) genes inRhizobium meliloti.J. Bacteriol. 149221.

    Google Scholar 

  • Ditta, G. (1986). Tn5 mapping ofRhizobium nitrogen fixation genes.Methods Enzymol. 118519.

    Google Scholar 

  • Durán, S., and Calderón, J. (1995). Role of the glutamine transaminase-θ-amidase pathway and glutaminase in glutamine degradation inRhizobium etli.Microbiology 141589.

    Google Scholar 

  • Durán, S., Du Pont, G., Huerta-Zepeda, A., and Calderón, J. (1995). The role of glutaminase inRhizobium etli: Studies with a new mutant.Microbiology 1412883.

    Google Scholar 

  • Espín, G., Moreno, S., and Guzman, J. (1994). Molecular genetics of the glutamine synthetases inRhizobium species.Crit. Rev. Microbiol. 20117.

    Google Scholar 

  • Friedman, A. M., Long, S. R., Brown, S. E., and Buikema, W. J. (1982). Construction of a broad host range cosmid cloning vector and its use in the genetic analysis ofRhizobium mutants.Gene 18289.

    Google Scholar 

  • Halpern, Y. S. (1988). Control of transport and utilization of nitrogen sources in bacteria. In Sanchez-Esquivel, S. (ed.),Nitrogen Source Control of Microbial Processes CRC Press, Boca Raton, FL, p. 21.

    Google Scholar 

  • Hartman, S. C. (1971). Glutaminase and γ-glutamyltransferases. In Boyer, P. D. (ed.),The Enzymes, Vol. 4 3rd edition, Academic Press, New York, pp. 79–100.

    Google Scholar 

  • Kahn, M. L., Kraus, J., and Somerville, J. E. (1985). A model of nutrient exchange in theRhizobium-legume symbiosis. In Evans, H., Bottomley, P., and Newton, W. E. (eds.), Nitrogen Fixation Research Progress, Nijhoff, New York, pp. 193–199.

    Google Scholar 

  • Katz, J., and Rognstad, R. (1976). Futiles cycles in the metabolism of glucose.Curr. Topics Cell. Regul. 10237.

    Google Scholar 

  • Kretovich, V. L., Sidel'nikova, L. I., Kasakova, O. V., Veinova, M. K., and Karyakina, T. I. (1981). Glutaminases and asparaginase of bacteroids fromRhizobium Lupini.Prikl. Biokhim. Mikrobio. 17821.

    Google Scholar 

  • Meade, H. M., Long, S. R., Ruvkun, G. B., Brown, S. E., and Ausubel, F. M. (1982). Physical and genetic characterization of symbiotic and auxotrophic mutants ofRhizobium meliloti in induced by transposon Tn5 mutagenesis.J. Bacteriol. 149114.

    Google Scholar 

  • Mora, J. (1990). Glutamine metabolism and cycling inNeurospora crassa.Microbiol. Rev. 54293.

    Google Scholar 

  • Osorio, A., Camarena, L., Salazar, G., Noll-Louzada, M., and Bastarrachea, F. (1993). Nitrogen regulation in anEscherichia coli strain with a temperature sensitive glutamyl-tRNA synthetase.Mol. Gen. Genet. 239400.

    Google Scholar 

  • Prusiner, S. (1973). Glutaminases ofEscherichia coli: Properties, regulation and evolution. In Prusiner, S., and Stadtman, E. R. (eds.),The Enzymes of Glutamine Metabolism Academic Press, New York, p. 293.

    Google Scholar 

  • Prusiner, S. (1975). Regulation of glutaminase levels inEscherichia coli.J. Bacteriol. 123992.

    Google Scholar 

  • Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989).Molecular Cloning: A Laboratory Manual 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

    Google Scholar 

  • Segovia, L., Young, P. W., and Martínez-Romero, E. (1993). Reclassification of AmericanRhizobium leguminosarum biovar phaseoli type I as Rhizobium etli sp.Nov. Int. J. Syst. Bacteriol. 43374.

    Google Scholar 

  • Smith, E. M., and Watford, M. (1990). Molecular Cloning of a cDNA for rat hepatic glutaminase.J. Biol. Chem. 26510631.

    Google Scholar 

  • Soberón, M., and González, A. (1987). Glutamine degradation through the ω-amidase pathway inSaccharomyces cerevisiae.J. Gen. Microbiol. 1339.

    Google Scholar 

  • Stadtman, E. R. (1973). A note on the significance of glutamine in intermediary metabolism. In Prusiner, S., and Stadtman, E. R. (eds.),The Enzymes of Glutamine Metabolism Academic Press, New York, p. 1.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Durán, S., Sánchez-Linares, L., Huerta-Saquero, A. et al. Identification of two glutaminases inRhizobium etli . Biochem Genet 34, 453–465 (1996). https://doi.org/10.1007/BF00570126

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00570126

Key words

Navigation