Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

The roles of nitrate and ammonium in the regulation of the development of nitrate reductase in Chlamydomonas reinhardii

Abstract

The regulation of the development of nitrate reductase (NR) activity in Chlamydomonas reinhardii has been compared in a wild-type strain and in a mutant (nit-A) which possesses a modified nitrate reductase enzyme that is non-functional in vivo. The modified enzyme cannot use NAD(P)H as an electron donor for nitrate reduction and it differs from wild-type enzyme in that NR activity is not inactivated in vitro by incubation with NAD(P)H and small quantities of cyanide; it is inactivated when reduced benzyl viologen or flavin mononucleotide is present. After short periods of nitrogen starvation mutant organisms contain much higher levels of terminal-NR activity than do similarly treated wild-type ones. Despite the inability of the mutant to utilize nitrate, no nitrate or nitrite was found in nitrogen-starved cultures; it is therefore concluded that the appearance of NR activity is not a consequence of nitrification. After prolonged nitrogen starvation (22 h) the NR level in the mutant is low. It increases rapidly if nitrate is then added and this increase in activity does not occur in the presence of ammonium, tungstate or cycloheximide. Disappearance of preformed NR activity is stimulated by addition of tungstate and even more by addition of ammonium. The results are interpreted as evidence for a continuous turnover of NR in cells of the mutant with ammonium both stimulating NR breakdown and stopping NR synthesis. Nitrate protects the enzyme from breakdown. Reversible inactivation of NR activity is thought to play an insignificant rôle in the mutant.

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

Abbreviations

NR:

nitrate reductase

BV:

benzyl viologen

References

  1. Beevers, L., Hageman, R.H. (1969) Nitrate reduction in higher plants. Ann. Rev. Plant Physiol. 19, 495–522

  2. Barea, J.L., Sosa, F., Cardenas, J. (1976) Cyanide inactivation of Chlamydomonas reinhardii nitrate reductase under reducing conditions. Z. Pflanzenphysiol. 79, 237–245

  3. Choudhary, V.P., Rao, G.R. (1976) Molecular basis of nitrate reductase induction in Candida utilis. Biochem. Biophys. Res. Commun. 72, 598–602

  4. Coddington, A. (1976) Biochemical studies on the nit mutant of Neurospora crassa. Mol. Gen. Genet. 145, 195–206

  5. Cove, D.J. (1970) Control of gene action in Aspergillus nidulans. Proc. Roy. Soc. (London) 176, 267–275

  6. Dantzig, A.H., Zurowski, W.K., Ball, T.M., Nason, A. (1978) Induction and repression of nitrate reductase in Neurospora crassa. J. Bacteriol. 133, 671–679

  7. Hewitt, E.J. (1975) Assimilatory nitrate-nitrite reduction. Ann. Rev. Plant Physiol. 26, 73–100

  8. Hipkin, C.R., Syrett, P.J. (1973) Enzymic determination of nitrate by use of frozen/thawed Chlorella cells. New Phytol. 72, 47–49

  9. Hipkin, C.R., Syrett, P.J. (1977a) Nitrate reduction by whole cells of Ankistrodesmus braunii and Chlamdomonas reinhardii. New Phytol 79, 639–648

  10. Hipkin, C.R., Syrett, P.J. (1977b) Post-transcriptional control of nitrate reductase formation in green algae. J. Exp. Bot. 28, 1270–1277

  11. Hipkin, C.R., Al-Bassam, B.A., Syrett, P.J. (1979a) Pyridine nucleotide specificity and other properties of purified nitrate reductase from Chlorella variegata. Planta 144, 137–141

  12. Hipkin, C.R., Syrett, P.J., Al-Bassam, B.A. (1979b) Some characteristics of nitrate reductase in unicellular algae. In: Nitrogen assimilation of plants, 6th Long Ashton Symposium, p. 309, Hewitt, E.J., Cutting, C.V., eds. Academic Press, London

  13. Kessler, E., Oesterheld, H. (1970) Nitrification and induction of nitrate reductase in nitrogen-deficient algae. Nature (London) 228, 287–288

  14. Losada, M. (1974) Interconversion of nitrate and nitrite reductase of the assimilatory type. In: Metabolic interconversions of enzymes. III Internat. Symp., p. 257, Fischer, E.H., Krebs, E.G., Neurath, H., Stadtman, E.R., eds. Springer, Berlin Heidelberg New York

  15. Losada, M., Herrera, J., Maldonado, J.M., Paneque, A. (1973) Mechanism of nitrate reductase reversible inactivation by ammonia in Chlamydomonas. Plant Sci. Lett. 1, 31–37

  16. Maldonado, J.M., Notton, B.A., Hewitt, E.J. (1978) Effects of reduced dehydrogenase electron acceptors on the various nitrate dependent activities of spinach (Spinacea oleracea L.) nitrate reductase. FEBS Lett. 93, 169–173

  17. Morris, I., Syrett, P.J. (1963) The development of nitrate reductase in Chlorella and its repression by ammonium. Arch. Mikrobiol. 47, 32–41

  18. Morton, A.J. (1955) A study of nitrate reduction in mould fungi. J. Exp. Bot. 7, 97–112

  19. Nichols, G.L., Syrett, P.J. (1978) Nitrate reductase deficient mutants of Chlamydomonas reinhardii. Isolation and genetics. J. Gen. Microbiol. 108, 71–77

  20. Nichols, G.L., Shehata, S.A.M., Syrett, P.J. (1978) Nitrate reductase deficient mutants of Chlamydomonas reinhardii. Biochemical characteristics. J. Gen. Microbiol. 108, 79–88

  21. Orebamjo, T.O., Stewart, G.R. (1974) Some characteristics of nitrate reductase induction in Lemna minor. Planta 117, 1–10

  22. Paneque, A., Vega, J.M., Cardenas, J., Herrera, J., Aparicio, P.J., Losada, M. (1972) 185Wo-labelled nitrate reductase from Chlorella. Plant Cell Physiol. 13, 175–178

  23. Premakumar, R., Sorger, G.J., Gooden, D. (1979) Nitrogen metabolite repression of nitrate reductase in Neurospora crassa. J. Bacteriol. 137, 1119–1126

  24. Solomonson, L.P. (1974) Regulation of nitrate reductase activity by NADH and cyanide. Biochim. Biophys. Acta 334, 297–308

  25. Solomonson, L.P., Spehar, A.M. (1977) Model for the regulation of nitrate assimilation. Nature (London) 265, 373–375

  26. Sorger, G.J., Davies, J. (1973) Regulation of nitrate reductase of Neurospora at the level of transcription and translation. Biochem. J. 134, 673–685

  27. Sorger, G.J., Debanne, M.T., Davies, J. (1974) Effects of nitrate on the synthesis and decay of nitrate reductase of Neurospora. Biochem. J. 140, 395–403

  28. Sorger, G.J., Premakumar, R., Gooden, D. (1978) Demonstration in vitro of two intracellular inactivators of nitrate reductase from Neurospora. Biochim. Biophys. Acta 540, 33–47

  29. Sosa, F.M., Cardenas, J. (1977) NADPH as electron donor for nitrate reduction in Chlamydomonas reinhardii. Z. Pflanzenphysiol 85, 171–175

  30. Spiller, H., Dietsch, E., Kessler, E. (1976) Intracellular appearance of nitrite and nitrate in nitrogen starved cells of Ankistrodesmus braunii. Planta 129, 175–181

  31. Stewart, G.R., Rhodes, D. (1977) Control of enzyme levels in the regulation of nitrogen assimilation. In: Regulation of enzyme synthesis and activity in higher plants, p. 1, Smith, H., ed. Academic Press, London New York San Francisco

  32. Subramanian, K.N., Sorger, G.J. (1972) Regulation of nitrate reductase in Neurospora crassa: Stability in vivo. J. Bacteriol. 110, 538–546

  33. Syrett, P.J., Hipkin, C.R. (1973) The appearance of nitrate reductase activity in nitrogen-starved cells of Ankistrodesmus braunii. Planta 111, 57–64

  34. Syrett, P.J., Leftley, J.W. (1976) Nitrate and urea assimilation by algae. In: Perspectives in experimental biology, p. 221, Sunderland, N., ed. Pergamon Press, Oxford New York

  35. Thomas, R.J., Hipkin, C.R., Syrett, P.J. (1976) The interaction of nitrogen assimilation with photosynthesis in nitrogen deficient cells of Chlorella. Planta 133, 9–13

  36. Wallace, W. (1975) Effects of a nitrate reductase inactivating enzyme and NAD(P)H on the nitrate reductase from higher plants and Neurospora. Biochim. Biophys. Acta 377, 239–250

  37. Zielke, R.H., Filner, P. (1971) Synthesis and turnover of nitrate reductase induced by nitrate in cultured tobacco cells. J. Biol. Chem. 146, 1772–1779

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Hipkin, C.R., Al-Bassam, B.A. & Syrett, P.J. The roles of nitrate and ammonium in the regulation of the development of nitrate reductase in Chlamydomonas reinhardii . Planta 150, 13–18 (1980). https://doi.org/10.1007/BF00385608

Download citation

Key words

  • Ammonium
  • Chlamydomonas (mutant)
  • Nitrate
  • Nitrate reductase