Skip to main content
Log in

Regulation of the nitrate-reducing system enzymes in wild-type and mutant strains of Chlamydomonas reinhardii

Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Six mutant strains (301, 102, 203, 104, 305, and 307) affected in their nitrate assimilation capability and their corresponding parental wild-type strains (6145c and 21gr) from Chlamydomonas reinhardii have been studied on different nitrogen sources with respect to NAD(P)H-nitrate reductase and its associated activities (NAD(P)H-cytochrome c reductase and reduced benzyl viologen-nitrate reductase) and to nitrite reductase activity. The mutant strains lack NAD(P)H-nitrate reductase activity in all the nitrogen sources. Mutants 301, 102, 104, and 307 have only NAD(P)H-cytochrome c reductase activity whereas mutant 305 solely has reduced benzyl viologen-nitrate reductase activity. Both activities are repressible by ammonia but, in contrast to the nitrate reductase complex of wild-type strains, require neither nitrate nor nitrite for their induction. Moreover, the enzyme from mutant 305 is always obtained in active form whereas nitrate reductase from wild-types needs to be reactivated previously with ferricyanide to be fully detected. Wild-type strains and mutants 301, 102, 104, and 307, when properly induced, exhibit an NAD(P)H-cytochrome c reductase distinguishable electrophoretically from contitutive diaphorases as a rapidly migrating band. Nitrite reductase from wild-type and mutant strains is also repressible by ammonia and does not require nitrate or nitrite for its synthesis. These facts are explained in terms of a regulation of nitrate reductase synthesis by the enzyme itself.

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.

Institutional subscriptions

Similar content being viewed by others

References

  • Bailey JL (1967) Techniques in protein chemistry, 2nd. Elsevier, Amsterdam, p 340

    Google Scholar 

  • Beevers L, Hageman RH (1969) Nitrate reduction in higher plants. Annu Rev Plant Physiol 20:495–522

    Google Scholar 

  • Coddington A (1976) Biochemical studies on the nit mutants of Neurospora crassa. Mol Gen Genet. 145:195–206

    Google Scholar 

  • Cove DJ, Pateman JA (1969) Autoregulation of the synthesis of nitrate reductase in Aspergillus nidulans. J Bacteriol 97:1374–1378

    Google Scholar 

  • Cullimore JV, Sims AP (1980) An association between photorespiration and protein catabolism: studies with Chlamydomonas. Planta 150:392–396

    Google Scholar 

  • Cullimore JV, Sims AP (1981) Glutamine synthetase of Chlamydomonas: its role in the control of nitrate assimilation. Planta 153:18–24

    Google Scholar 

  • Fernández E (1981) Caracterización molecular del complejo enzimático NAD(P)H-nitrato reductase de algas verdes mediante el estudio de estirpes silvestres y mutantes de Chlamydomonas reinhardii. Doctoral Dissertation, University of Sevilla, Spain

    Google Scholar 

  • Fernández E, Cárdenas J (1981a) In vitro complementation of assimilatory NAD(P)H-nitrate reductase from mutants of Chlamydomonas reinhardii. Biochim Biophys Acta 657:1–12

    Google Scholar 

  • Fernández E, Cárdenas J (1981b) Occurrence of xanthine dehydrogenase in Chlamydomonas reinhardii: A common cofactor shared by xanthine dehydrogenase and nitrate reductase. Planta 153:254–257

    Google Scholar 

  • Florencio FJ, Vega JM (1981) Regulación de la asimilación de nitrato en Chlamydomonas Chlamydomonas reinhardii. Proceedings of the Second Congress of the Spanish Fed of Exp Biol Soc. FESBE, Madrid, p 96

  • Florencio FJ, Vega JM, Losada M (1980) Efecto del amonio y de la obscuridad en la asimilación fotosintética del nitrato por el alga verde Chlamydomonas reinhardii. Proceedings of the First Spanish-Portuguese Congress of Biochem. SEB-SPB, Coimbra p 195

  • Flores E, Guerrero MG Losada M (1980) Short-term ammonium inhibition of nitrate utilization by Anacystis nidulans and other cyanobacteria. Arch Microbiol 128:137–144

    Google Scholar 

  • Garrett RH, Amy NK (1978) Nitrate assimilation in fungi. Adv Microbiol Physiol 18:1–65

    Google Scholar 

  • Herrera J, Paneque A, Maldonado JM, Barea JL, Losada M, (1972) Regulation by ammonia of nitrate reductase synthesis and activity in Chlamydomonas reinhardii. Biochem Biophys Res Commun 48:996–1003

    Google Scholar 

  • Hipkin CR, Al-Bassam BA, Syrett PJ (1980) The roles of nitrate and ammonium in the regulation of the development of nitrate reductase in Chlamydomonas reinhardii. Planta 150:13–18

    Google Scholar 

  • Jovin T, Charambach A, Naughton MA (1964) Apparatus for preparative temperature regulated polyacrylamide gel electrophoresis. Anal Biochem 9:351–364

    Google Scholar 

  • Kaplan, D, Mayer AM, Lips SH (1978) Nitrite activation of nitrate reductase in higher plants. Planta 138:205–209

    Google Scholar 

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

    Google Scholar 

  • Losada M, Guerrero MG, Vega JM (1981) The assimilatory reduction of nitrate. In: Bothe H, Trebst A (eds) Biology on inorganic nitrogen and sulfur. Springer, Berlin Heidelberg New York, pp 32–63

    Google Scholar 

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

    Google Scholar 

  • Losada M, Paneque A, Aparicio PJ, Vega JM, Cárdenas J, Herrera J (1970) Inactivation and repression by ammonium of the nitrate reducing system in Chlorella. Biochem Biophys Res Commun 38:1009–1015

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

  • MacDonald DW, Cove DJ, Coddington A (1974) Cytochrome c reductases from wild-type and mutant strains of Aspergillus nidulans. Mol Gen Genet 128:187–199

    Google Scholar 

  • Maldonado JM, James DM, Notton BA, Hewitt EJ (1980) Occurrence and properties of cytochrome c reductase in cereals. Proceedings of the Second Congress of the Fed of Eur Soc of Plant Physiol. FESPP Santiago, pp 468–469

    Google Scholar 

  • Mendel RR Müller AJ (1979) Nitrate reductase-deficient mutant cell lines of Nicotiana tabacum. Further biochemical characterization. Mol Gen Genet 177:145–153

    Google Scholar 

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

    Google Scholar 

  • Morris GJ, Coulson G, Clarke A (1979) The cryopreservation of Chlamydomonas. Cryobiology 16:401–410

    Google Scholar 

  • Nichols GL, Shehata SAM, Syrett PJ (1978) Nitrate reductase deficient mutants of Chlamydomonas reinhardii. Biochemical characteristics. J Gen Microbiol 108:79–88

    Google Scholar 

  • Palacian E, De la Rosa FF, Castillo F, Gomez-Moreno C (1974) Nitrate reductase from Spinacea oleracea. Reversible inactivation by NAD(P)H and by thiols. Arch Biochem Biophys 161:441–447

    Google Scholar 

  • Paneque A, Losada M (1966) Comparative reduction of nitrate by spinach nitrate reductase with NADH2 and NADPH2. Biochim Biophys Acta 128:202–204

    Google Scholar 

  • Paneque A, Del Campo FF, Ramirez JM, Losada M (1965) Flavin nucleotide nitrate reductase from spinach. Biochim Biophys Acta 109:79–85

    Google Scholar 

  • Pistorius EK, Fünkhouser EA, Voss H (1978) Effect of ammonium and ferricyanide on nitrate utilization by Chlorella vulgaris. Planta 141:279–282

    Google Scholar 

  • Ramirez JM, Del Campo FF, Paneque A, Losada M (1966) Ferredoxin-nitrite reductase from spinach. Biochim Biophys Acta 118:58–71

    Google Scholar 

  • Snell FD, Snell CT (1949) Colorimetric methods of analysis, vol 2. Van Nostrand, New York, p 804

    Google Scholar 

  • Sosa FM, Ortega T, Barea JL (1978) Mutants from Chlamydomonas reinhardii affected in their nitrate assimilation capabiligy. Plant Sci Lett 11:51–58

    Google Scholar 

  • Subramanian KN, Sorger GJ (1972) The role of molybdenum in the synthesis of Neurospora nitrate reductase. Biochim Biophys Acta 256:533–543

    Google Scholar 

  • Thacker A, Syrett PJ (1972) Disappearance of nitrate reductase activity from Chlamydomonas reinhardii. New Phytol 71:435–441

    Google Scholar 

  • Tomsett AB, Garrett RH (1981) Biochemical analysis of mutants defective in nitrate assimilation in Neurospora crassa: evidence for autogenous control by nitrate reductase. Mol Gen Genet 184:183–190

    Google Scholar 

  • Vennesland B, Guerrero MG (1979) Reduction of nitrate and nitrite. In: Gibbs M, Latzko E (eds) Encyclopedia of plant physiol, New Series, vol 6 Springer-Verlag, Berlin, pp 425–444

    Google Scholar 

  • Wallace W, Johnson CB (1978) Nitrate reductase and soluble cytochrome c reductase(s) in higher plants. Plant Physiol 61:748–752

    Google Scholar 

  • Wang CC, Raper JR (1970) Isozyme patterns and sexual morphogenesis in Schizophyllum. Proc Natl Acad Sci USA 66:882–889

    Google Scholar 

  • Wray JL, Filner P (1970) Structural and functional relationships of enzyme activities induced by nitrate in barley. Biochem J 119:715–725

    Google Scholar 

  • Zumft WG, Paneque A, Aparicio PJ, Losada M (1969) Mechanism of nitrate reduction in Chlorella. Biochem Biophys Res Commun 36:980–986

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by H. Böhme

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fernández, E., Cárdenas, J. Regulation of the nitrate-reducing system enzymes in wild-type and mutant strains of Chlamydomonas reinhardii . Mol Gen Genet 186, 164–169 (1982). https://doi.org/10.1007/BF00331846

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00331846

Keywords

Navigation