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Effect of glutamine on glutamine-synthetase regulation in the green alga Monoraphidium braunii

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Abstract

Glutamine-synthetase (GS; EC 6.3.1.2) activity and protein levels were measured in crude extracts from Monoraphidium braunii Näegeli, strain 202-7d, cultures grown under different nitrogen sources. Only ammonium and l-glutamine promoted a partial enzyme inactivation, which, in the case of l-glutamine, was accompanied by a significant repression of GS. Methionine sulfoximine (MSX), a strong inhibitor of GS, produced a drastic inactivation of GS which was concomitant with a marked increase in GS protein as measured by rocket immunoelectrophoresis. Such an increase was prevented in the presence of cycloheximide. The effect of the l-glutamine analog on GS activity and protein was partially inhibited if l-glutamine was also added to cell cultures, possibly indicating competition in the transport of these two substances. In addition, the effects of MSX were reversed after longer times when cultures were treated with smaller concentrations of inhibitor. Treatment of cell cultures with azaserine, a specific inhibitor of glutamate synthase, the second enzyme acting in the ammonium assimilation pathway, promoted a strong GS inactivation and a partial repression of this enzyme, which paralleled a specific increase in the intracellular pools of glutamine High-performance liquid chromatography measurements of intracellular amino-acid concentrations showed that glutamine levels correlated negatively with GS concentration. A role for glutamine as a negative effector of GS synthesis is proposed.

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Abbreviations

GS:

l-glutamine synthetase

GOGAT:

l-glu-tamine:2-oxoglutarate amidotransferase

MSX:

methionine sulfoximine

References

  • Ahmad, I., Hellebust, J.A. (1984) Nitrogen metabolism of the marine microalga Chlorella autotrophica. Plant Physiol. 76, 658–663

    Google Scholar 

  • Ahmad, I., Hellebust, J.A. (1987) Glutamine synthetase isoenzymes in the green soil alga Stichococcus bacillaris Naeg. Plant Physiol. 83, 259–261

    Google Scholar 

  • Ahmad, I., Hellebust, J.A. (1990) Regulation of chloroplast development by nitrogen source and growth conditions in a Chlorella protothecoides strain. Plant Physiol. 94, 944–949

    Google Scholar 

  • Alef, K., Zumft, W.G. (1981) Regulatory properties of glutamine synthetase from the nitrogen fixing phototrophic bacterium Rhodopseudomonas palustris. Z. Naturforsch. 36c, 784–789

    Google Scholar 

  • Alef, K., Burkhardt, HJ., Horstmann, H.J., Zumft, W.G. (1981) Molecular characterization of glutamine synthetase from the nitrogen fixing phototrophic bacterium Rhodopseudomonas palustris. Z. Naturforsch. 36c, 246–254

    Google Scholar 

  • Akimova, N.I., Evstigneeva, Z.G., Kretovich, V.L. (1976) Regulation of the glutamine metabolism in Chlorella pyrenoidosa. Investigation of the mechanisms of the regulation of glutamine synthetase activity in the process of assimilation of ammonium. Biochemistry (U.S.S.R.) 41, 1306–1312

    Google Scholar 

  • Ayling, P.D. (1981) Methionine sulfoximine is transported by high affinity methionine and glutamine transport system in Salmonella typhimurium. J. Bacteriol. 148, 514–520

    Google Scholar 

  • Bailey, J.L. (1967) Techniques in protein chemistry, 2nd edn., pp. 340–341, Elsevier Science Publishing Inc., New York

    Google Scholar 

  • Beudeker, R.F., Tabita, F.R. (1985) Characterization of glutamine synthetase isoforms from Chlorella. Plant Physiol. 77, 791–794

    Google Scholar 

  • Carnicero, A., Mansito, T.B., Roldán, J.M., Falcón, M.A. (1990) Staphylolytic enzyme from Pseudomonas aeruginosa: characterization and immunocytochemical localization. Arch. Microbiol. 154, 37–41

    Google Scholar 

  • Casselton, P.J., Chandler, G., Shah, N., Stewart, G.R., Sumar, N. (1986) glutamine synthetase isoforms in algae. New Phytol. 102, 261–270

    Google Scholar 

  • Chapman, J.S., Meeks, J.C (1983) glutamine and glutamate transport by Anabaena variabilis. J. Bacteriol. 156, 122–129

    Google Scholar 

  • Cullimore, J.V., Sims, A.P. (1981a) glutamine synthetase of Chlamydomonas: its role in the control of nitrate assimilation. Planta 153, 18–24

    Google Scholar 

  • Cullimore, J.V., Sims, A.P. (1981b) Pathway of ammonium assimilation in illuminated and darkened Chlamydomonas reinhardtii. Phytochemistry 20, 933–940

    Google Scholar 

  • Diez, J., López-Ruiz, A. (1989) Immunological approach to the regulation of nitrate reductase in Monoraphidium braunii. Arch. Biochem. Biophys. 268, 707–715

    Google Scholar 

  • Engelhardt, H., Klemme, J.H. (1981) In vivo control of glutamine synthetase in the facultative phototrophic bacterium Rhodopseudomonas spheroides. Z. Naturforsch. 36c, 407–410

    Google Scholar 

  • Feng, B., Shiber, S.K., Max, S.R. (1990) Glutamine regulates glutamine synthetase expression in skeletal muscle cells in culture. J. Cell. Physiol. 145, 376–380

    Google Scholar 

  • Florencio, F.J., Vega, J.M. (1983) Separation, purification, and characterization of two isoforms of glutamine synthetase from Chlamydomonas reinhardtii. Z. Naturforsch. 38c, 531–538

    Google Scholar 

  • García-Fernández, J.M., López-Ruiz, A., Toribio, F., Roldán, J.M., Diez, J. (1994) Occurrence of only one form of glutamine synthetase in the green algae Monoraphidium braunii. Plant Physiol. 104, 425–430

    Google Scholar 

  • Johansson, B.C., Gest, H. (1977) Adenylylation/deadenylylation control of the glutamine synthetase of Rhodopseudomonas capsulata. Eur. J. Biochem. 81, 365–371

    Google Scholar 

  • Kleiner, D., Alef, K., Hartmann, A. (1983) Uptake of methionine sulfoximine by some N2-fixing bacteria, and its effect on ammonium transport. FEBS Lett. 164, 121–123

    Google Scholar 

  • Menacho, A., Vega, J.M. (1989) Effect of nitrogen starvation on ammonium assimilation by Chlamydomonas reinhardtii. Physiol. Plant. 75, 285–289

    Google Scholar 

  • Mérida, A., Leurentop, L., Candau, P., Florencio, F.J. (1990) Purification and properties of glutamine synthetase from the cyanobacteria Synechocystis sp strain PCC 6803 and Calothrix sp strain PCC 7601. J. Bacteriol. 172, 4732–4735

    CAS  PubMed  Google Scholar 

  • Mérida, A., Candau, P., Florencio, F.J. (1991) Regulation of glutamine synthetase activity in the unicellular cyanobacterium Synechocistis sp strain PCC 6803 by the nitrogen source: effect of ammonium. J Bacteriol. 173, 4095–4100

    Google Scholar 

  • Moreno-Vivián, C., Cejudo, F.J., Gárdenas, J., Castillo, F. (1983) Ammonium assimilation pathways in Rhodopseudomonas capsulata E1F1. Arch. Microbiol. 136, 147–151

    Google Scholar 

  • Nagatani, H., Shimizu, M., Valentine, R.C. (1971) The mechanism of ammonium assimilation in nitrogen fixing bacteria. Arch. Microbiol. 79, 164–175

    Google Scholar 

  • Nordlund, S., Kanemoto, R.H., Murrell, S.A., Ludden, P.W. (1985) Properties and regulation of glutamine synthetase from Rhodospirillum rubrum. J. Bacteriol. 161, 13–17

    Google Scholar 

  • Orr, J., Haselkorn, R. (1982) Regulation of glutamine synthetase activity and synthesis in free-living and symbiotic Anabaena sp. J. Bacteriol. 152, 626–635

    Google Scholar 

  • Paul, J.H., Cooksey, K.E. (1981) Regulation of asparaginase, glutamine synthetase and glutamate dehydrogenase in response to medium nitrogen concentrations in a euryhaline Chlamydomonas species. Plant Physiol. 68, 1364–1368

    Google Scholar 

  • Rasulov, A.S., Shakirov, Z.S., Evstigneeva, Z.G., Tsuprun, V.L., Kretovich, V.L. (1986) Quaternary structure of glutamine synthetase of Ankistrodesmus braunii. Biochemistry (USSR) 51, 342–347

    Google Scholar 

  • Rasulov, A.S., Shakirov, Z.S., Evstigneeva, Z.G., Kretovich, V.L. (1989) Adenylated control of the activity of molecular forms of glutamine synthetase from Ankistrodesmus braunii. Biochemistry (USSR) 54, 1664–1667

    Google Scholar 

  • Romero, F., Caballero, F.J., Castillo, F., Roldán, J.M. (1985) Immunoelectrophoretic approach to the metabolic regulation of glutamine synthetase in Rhodopseudomonas capsulata E1F1:role of glutamine. Arch. Microbiol. 143, 111–116

    Google Scholar 

  • Romero, F., Quintero, A., Roldán, J.M. (1989a) Role of glutamine as a direct co-repressor of glutamine synthetase in Rhodobacter capsulatus E1F1. FEMS Microbiol. Lett. 58, 111–114

    Google Scholar 

  • Romero, F., Quintero, A., Roldán, J.M. (1989b) Characterization of a methionine sulfoximine resistant strain of Rhodobacter capsulatus E1F1. FEMS Microbiol. Lett. 61, 203–206

    Google Scholar 

  • Scmidt, A. (1981) A thioredoxin activated glutamine synthetase in Chlorella. Z. Naturforsch. 36c, 396–399

    Google Scholar 

  • Shpiro, B.M., Stadtman, E.R. (1970) glutamine synthetase (Escherichia coli). Methods Enzymol 17A, 910–922

    Google Scholar 

  • Stacey, G., Van Baalen, C., Tabita, R. (1979) Nitrogen and ammonium assimilation in the cyanobacteria. Regulation of glutamine synthetase. Arch. Biochem. Biophys. 206, 181–189

    Google Scholar 

  • Stedtman, E.R., Ginsburg, A., Ciardi, J.E., Yeh, J., Henning, S.B., Shapiro, B.M. (1970) Multiple molecular forms of glutamine synthetase produced by enzyme catalyzed adenylylation and deadenylylation reactions. Adv. Enzyme Reg. 8, 99–118

    Google Scholar 

  • Stedtman, E.R., Ginsburg, A. (1974) The glutamine synthetase of Escherichia coli, structure and control. In: The enzymes, vol. 10, pp. 755–807, Boyer, P., ed. Academic Press Inc, New York

    Google Scholar 

  • Sumar, N., Casselton, P.J., McNally, S.F., Stewart, G.R. (1984) Occurrence of isoenzymes of glutamine synthetase in the alga Chlorella kessleri. Plant Physiol. 74, 204–207

    Google Scholar 

  • Tuli, R., Thomas, J. (1980) Regulation of glutamine synthetase in the blue-green alga Anabaena L-31. Biochim. Biophys. Acta 613, 526–533

    Google Scholar 

  • Tuli, R, Thomas, J. (1981) In vivo regulation of glutamine synthetase by ammonium in the cyanobacterium Anabaena L-31. Arch. Biochem. Biophys. 206, 181–189

    Google Scholar 

  • Ullrich, W.R., Aparicio, P.J., Syrett, P.J., Castillo, F., eds (1987) Inorganic nitrogen metabolism. Springer, Berlin

    Google Scholar 

  • Vendrell, J., Avilés, F.X. (1986) Complete amino-acid analysis of protein by dabsyl derivatization and reversed-phase liquid chromatography. J. Chromatogr. 358, 401–413

    Google Scholar 

  • Weeke, B. (1973) Rocket immunoelectrophoresis. In: A manual of quantitative immunolectrophoresis. Methods and applications, pp 37–46, Axelsen, N.H., Krøll, J., Weeke, B., eds. Universitetsforlaget, Oslo

    Google Scholar 

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Authors and Affiliations

  1. Departamento de Bioquímica y Biología Molecular, Instituto de Biología Básica y Aplicada, Facultad de Veterinaria, Universidad de Córdoba, Avda. Medina Azahara, s/n, E-14071, Córdoba, Spain

    José Manuel Garcia-Fernández, Antonio López-Ruiz, José Alhama, José Manuel Roldán & Jesús Diez Dapena

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  1. José Manuel Garcia-Fernández
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  2. Antonio López-Ruiz
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  3. José Alhama
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  4. José Manuel Roldán
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  5. Jesús Diez Dapena
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Correspondence to Jesús Diez Dapena.

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During the course of this work, J.A. was supported by a fellowship from Junta de Andalucía, and J.M. G-F. by a fellowship from the Spanish Ministerio de Educatión y Ciencia. This work was supported by the Junta de Andalucía.

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Garcia-Fernández, J.M., López-Ruiz, A., Alhama, J. et al. Effect of glutamine on glutamine-synthetase regulation in the green alga Monoraphidium braunii . Planta 195, 434–439 (1995). https://doi.org/10.1007/BF00202602

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