Skip to main content
SpringerLink
Log in

Molecular Control of Nitrate Reductase and Other Enzymes Involved in Nitrate Assimilation

  • Chapter
Photosynthetic Nitrogen Assimilation and Associated Carbon and Respiratory Metabolism

Part of the book series: Advances in Photosynthesis and Respiration ((AIPH,volume 12))

Summary

Nitrate acts as both a nutrient and a signal in plants. Nitrate induces gene expression of enzymes for its metabolism into amino acids but also has other effects on plant metabolism and development. Familiar nitrate-induced enzymes are nitrate and nitrite reductases, nitrate transporters, glutamine synthetase, glutamate synthase, ferredoxin and ferredoxin NADP+ reductase. Microarray analysis of nitrate-stimulated gene expression has identified 40 transcripts including hemoglobin, transaldolase, regulatory and stress proteins, several protein kinases and several methyltransferases. Coordinated expression of these nitrate-stimulated genes is probably due to a single ‘nitrate-transacting factor’ and a ‘nitrate’ box has been elucidated for nitrate and nitrite reductase genes with constitutively expressed nuclear proteins which bind to the box. A MADS transcription factor is nitrate-induced in roots and involved in development of lateral roots. However, accumulation of nitrate overcomes this signal and halts lateral root development. Post-translational inhibition of nitrate reductase activity illustrates a complex control mechanism involving protein phosphorylation and binding of the ubiquitous binding protein called 14-3-3. Protein kinases catalyzing phosphorylation have been identified and 14-3-3 binding elucidated, which includes activation of 14-3-3 by polycations such as polyamines. Using molecular modeling, it was shown that one 14-3-3 binding site can bind to the nitrate reductase dimer. Nitrate reductase-14-3-3 complexes could bind via the second binding site on 14-3-3 to another enzyme/protein with a 14-3-3 binding site or nitrate reductase aggregation could result in rapid degradation. Two types of protein kinases are involved in nitrate reductase phosphorylation: calcium-dependent protein kinases and SnRKs (enzymes related to yeast sucrose non-fermenting (SNF1) protein kinases). Calcium-dependent protein kinases are activated by environmental and development signals via changes in intracellular calcium level, which impacts many plant metabolic pathways. SnRKs are less well understood and may be responding to more general metabolic signals.

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

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 219.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Athwal GS, Huber JL and Huber SC (1998) Phosphorylated nitrate reductase and 14-3-3 proteins. Site of interaction, effects of ions, and evidence for an AMP-binding site on 14-3-3 proteins. Plant Physiol 118: 1041–1048

    Article  CAS  PubMed  Google Scholar 

  • Athwal GS, Lombardo CR, Huber JL, Masters SC, Fu H and Huber SC (2000) Modulation of 14-3-3 protein interactions with target peptides by physical and metabolic effectors. Plant Cell Physiol 41: 523–533

    CAS  PubMed  Google Scholar 

  • Bachmann M, Shiraishi N, Campbell WH, Yoo BC, Harmon AC and Huber SC (1996a) Identification of Ser-543 as the major regulatory phosphorylation site in spinach leaf nitrate reductase. Plant Cell 8: 505–517

    Article  CAS  PubMed  Google Scholar 

  • Bachmann M, Huber JL, Liao PC, Gage DA and Huber SC (1996b) The inhibitor protein of phosphorylated nitrate reductase from spinach (Spinacia oleracea) leaves is a 14-3-3 protein. FEBS Lett 387: 127–131

    Article  CAS  PubMed  Google Scholar 

  • Bachmann M, Huber JL, Athwal GS, Wu K, Ferl RJ and Huber SC (1998) 14-3-3 proteins associate with the regulatory phosphorylation site of spinach leaf nitrate reductase in an isoform-specific manner and reduce dephosphorylation of Ser-543 by endogenous protein phosphatases. FEBS Lett 398: 26–30

    Google Scholar 

  • Campbell WH (1988) Nitrate reductase and its role in nitrate assimilation in plants. Physiol Plant 74: 214–219

    CAS  Google Scholar 

  • Campbell WH (1996) Nitrate reductase biochemistry comes of age. Plant Physiol 111: 355–361

    CAS  PubMed  Google Scholar 

  • Campbell WH (1999) Nitrate reductase structure, function and regulation: Bridging the gap between biochemistry and physiology. Annu Rev Plant Physiol Plant Mol Biol 50: 277–303

    Article  CAS  PubMed  Google Scholar 

  • Campbell WH and Kinghorn JR (1990) Functional domains of assimilatory nitrate reductases and nitrite reductases. Trends Biochem Sci 15: 315–319

    Article  CAS  PubMed  Google Scholar 

  • Chandok MR and Sopory SK (1996) Phosphorylation/dephosphorylation steps are key events in the phytochrome-mediated enhancement of nitrate reductase mRNA levels and enzyme activity in maize. Mol Gen Genet 251: 599–608

    CAS  PubMed  Google Scholar 

  • Cheng CL, Acedo GN, Cristinsin M and Conkling MA (1992) Sucrose mimics the light induction of Arabidopsis nitrate reductase gene transcription. Proc Natl Acad Sci USA 89: 1861–1864

    CAS  PubMed  Google Scholar 

  • Cotelle V, Meek SE, Provan F, Milne FC, Morrice N and MacKintosh C (2000) 14-3-3s regulate global cleavage of their diverse binding partners in sugar-starved Arabidopsis cells. EMBO J 19: 2869–2876

    Article  CAS  PubMed  Google Scholar 

  • Crawford N (1995) Nitrate: Nutrient and signal for plant growth. Plant Cell 7: 859–868

    Article  CAS  PubMed  Google Scholar 

  • Douglas P, Morrice N and MacKintosh C (1995) Identification of a regulatory phosphorylation site in the hinge 1 region of nitrate reductase from spinach (Spinacea oleracea) leaves. FEBS Lett 377: 113–117

    Article  CAS  PubMed  Google Scholar 

  • Douglas P, Moorhead G, Hong Y, Morrice N and MacKintosh C (1998) Purification of a nitrate reductase kinase from Spinacea oleracea leaves, and its identification as a calmodulin-domain protein kinase. Planta 206: 435–442

    Article  CAS  PubMed  Google Scholar 

  • Durner J and Klessig DF (1999) Nitric oxide as a signal in plants. Curr Opin Plant Biol 2: 369–374

    Article  CAS  PubMed  Google Scholar 

  • Dwivedi UP, Shiraishi N and Campbell WH (1994) Identification of an ‘essential’ cysteine of nitrate reductase via mutagenesis of its recombinant cytochrome b reductase domain. J Biol Chem 269: 13785–13791

    CAS  PubMed  Google Scholar 

  • Forde BG (2000) Nitrate transporters in plants: Structure, function and regulation. Biochim Biophys Acta 1465: 219–235

    CAS  PubMed  Google Scholar 

  • George GN, Mertens JA and Campbell WH (1999) Structural changes induced by catalytic turnover at the molybdenum site of Arabidopsis nitrate reductase. J Am Chem Soc 121: 9730–9731

    CAS  Google Scholar 

  • Gowri G, Ingemarsson B, Redinbaugh M and Campbell WH (1992) Nitrate reductase transcript is expressed in the primary response of maize to environmental nitrate. Plant Mol Biol 18: 55–64

    Article  CAS  PubMed  Google Scholar 

  • Halford NG and Hardie DG (1998) SNF1-related protein kinases: Global regulators of carbon metabolism in plants? Plant Mol Biol 37: 735–748

    Article  CAS  PubMed  Google Scholar 

  • Hardie DG, Carling D and Carlson M (1998) The AMP-activated/SNF1 protein kinase subfamily: Metabolic sensors of the eukaryotic cell? Annu Rev Biochem 67: 821–855

    Article  CAS  PubMed  Google Scholar 

  • Harmon AC, Gribskov M and Harper JF (2000) CDPKs—a kinase for every Ca2+ signal? Trends Plant Sci 5: 154–159

    Article  CAS  PubMed  Google Scholar 

  • Huber JL, Huber SC, Campbell WH and Redinbaugh MG (1992) Reversible light/dark modulation of spinach leaf nitrate reductase activity involves protein phosphorylation. Arch Biochem Biophys 296: 58–65

    Article  CAS  PubMed  Google Scholar 

  • Huber SC, Bachmann M and Huber JL (1996) Post-translational regulation of nitrate reductase activity: A role for Ca2+ and 14-3-3 proteins. Trends Plant Sci 1: 432–438

    Article  Google Scholar 

  • Hwang CF, Lin Y, D’Souza T and Cheng CL (1997) Sequences necessary for nitrate-dependent transcription of Arabidopsis nitrate reductase genes. Plant Physiol 113: 853–862

    Article  CAS  PubMed  Google Scholar 

  • Kaiser WM and Spill D (1991) Rapid modulation of spinach leaf nitrate reductase by photosynthesis. II. In vitro modulation by ATP and AMP. Plant Physiol 96: 368–375

    CAS  Google Scholar 

  • Kanamaru K, Wang R, Su W and Crawford NM (1999) Ser-534 in the hinge 1 region of Arabidopsis nitrate reductase is conditionally required for binding of 14-3-3 proteins and in vitro inhibition. J Biol Chem 274: 4160–4165

    Article  CAS  PubMed  Google Scholar 

  • Kaye C, Crawford NM and Malmberg RL (1997) Constitutive non-inducible expression of the Arabidopsis thaliana Nia 2 gene in two nitrate reductase mutants of Nicotiana plumbaginifolia. Plant Mol Biol 33: 953–964

    Article  CAS  PubMed  Google Scholar 

  • Kisker C, Schindelin H, Pacheco A, Wehbi WA, Garrett RM, Rajagopalan KV, Enemark JH and Rees DC (1997) Molecular basis of sulfite oxidase deficiency from the structure of sulfite oxidase. Cell 91: 973–983

    Article  CAS  PubMed  Google Scholar 

  • Lin Y, Hwang CF, Brown JB and Cheng CL (1994) 5′ proximal regions of Arabidopsis nitrate reductase genes direct nitrate-induced transcription in transgenic tobacco. Plant Physiol 106: 477–484

    Article  CAS  PubMed  Google Scholar 

  • Lu G, Campbell WH, Schneider G and Lindqvist Y (1994) Crystal structure of the FAD-containing fragment of corn nitrate reductase at 2.5 Å resolution: Relationship to other flavoprotein reductases. Structure 2: 809–821

    Article  CAS  PubMed  Google Scholar 

  • Lu G., Lindqvist Y, Schneider G, Dwivedi UN and Campbell WH (1995) Structural studies on corn nitrate reductase. Refined structure of the cytochrome b reductase fragment at 2.5 Å, its ADP complex and an active site mutant and modeling of the cytochrome b domain. J Mol Biol 248: 931–948

    Article  PubMed  Google Scholar 

  • MacKintosh C (1992) Regulation of spinach-leaf nitrate reductase by reversible phosphorylation. Biochim Biophys Acta 1137: 121–126

    CAS  PubMed  Google Scholar 

  • MacKintosh C (1998) Regulation of cytosolic enzymes in primary metabolism by reversible protein phosphorylation. Curr Opin Plant Biol 1: 224–229

    Article  CAS  PubMed  Google Scholar 

  • Matsumara T, Sakakibara H, Nakano R, Kimata Y, Sugiyama T and Hase T (1997) A nitrate-inducible ferredoxin in maize roots: Genomic organization and differential expression of two non-photosynthetic ferredoxin apoproteins. Plant Physiol 114: 653–660

    Google Scholar 

  • McMichael RW, Bachmann M and Huber SC (1995) Spinach leaf sucrose-phosphate synthase and nitrate reductase are phosphorylated/inactivated by multiple protein kinases in vitro. Plant Physiol 108: 1077–1082

    CAS  PubMed  Google Scholar 

  • Mendel RR (1997) Molybdenum cofactor of higher plants: Biosynthesis and molecular biology. Planta 203: 399–405

    Article  CAS  PubMed  Google Scholar 

  • Mertens JA, Campbell WH, Skipper L and Lowe DJ (1999) Electron transfer from FAD to heme-Fe in plant NADH: nitrate reductase. In: Ghisla S, Kroneck P, Macheroux P and Sund H (eds) Flavins and Flavoproteins 1999, pp 131–134. Agency for Scientific Publications, Berlin

    Google Scholar 

  • Mertens JA, Shiraishi N and Campbell WH (2000) Recombinant expression of molybdenum reductase fragments of plant nitrate reductase at high levels in Pichia pastoris. Plant Physiol 123: 743–756

    Article  CAS  PubMed  Google Scholar 

  • Meyer C, Levin JM, Roussel J-M and Rouze P (1991) Mutational and structural analysis of the nitrate reductase heme domain of Nicotiana plumbaginifolia. J Biol Chem 266: 20561–20566

    CAS  PubMed  Google Scholar 

  • Moorhead G, Douglas P, Morrice N, Scarabel M, Aitken A and MacKintosh C (1996) Phosphorylated nitrate reductase from spinach leaves is inhibited by 14-3-3 proteins and activated by fusicoccin. Curr Biol 6: 1104–1113

    Article  CAS  PubMed  Google Scholar 

  • Moorhead G, Douglas P, Cotelle V, Harthill J, Morrice N, Meek S, Deiting U, Stitt M, Scarabel M, Aitken A and MacKintosh C (1999) Phosphorylation-dependent interactions between enzymes of plant metabolism and 14-3-3 proteins. Plant J 18: 1–12

    Article  CAS  PubMed  Google Scholar 

  • National Research Council (2000) Clean Coastal Waters: Understanding and Reducing the Effects of Nutrient Pollution. Committee on the Causes and Management of Eutrophication, National Research Council, National Academy Press, Washington, DC

    Google Scholar 

  • Petosa C, Masters SC, Bankston LA, Pohl J, Wang B, Fu H and Liddington RC (1998) 14-3-3zeta binds a phosphorylated raf peptide and an unphosphorylated peptide via its conserved amphipathic groove. J Biol Chem 273: 16305–16310

    Article  CAS  PubMed  Google Scholar 

  • Pigaglio E, Durand N and Meyer C (1999) A conserved acidic motif in the N-terminal domain of nitrate reductase is necessary for the inactivation of the enzyme in the dark by phosphorylation and 14-3-3 binding. Plant Physiol 119: 219–230

    Article  CAS  PubMed  Google Scholar 

  • Provan F, Aksland L-M, Meyer C and Lillo C (2000) Deletion of the nitrate reductase N-terminal domain still allows binding of 14-3-3 proteins but affects their inhibitory properties. Plant Physiol 123: 757–764

    Article  CAS  PubMed  Google Scholar 

  • Rastogi R, Back E, Schneiderbauer A, Bowsher CG, Moffat B and Rothstein SJ (1993) A 330 bp region of the spinach nitrite reductase gene promoter directs nitrate-inducible tissue-specific expression in transgenic tobacco. Plant J 4: 317–326

    Article  CAS  Google Scholar 

  • Rastogi R, Bate NJ, Sivasankar S and Rothstein SJ (1997) Footprinting of the spinach nitrite reductase gene promoter reveals the preservation of nitrate regulatory elements between fungi and higher plants. Plant Mol Biol 34: 465–476

    Article  CAS  PubMed  Google Scholar 

  • Ratnam K, Shiraishi N, Campbell WH and Hille R (1995) Spectroscopic and kinetic characterization of the recombinant wild-type and C242S mutant of the cytochrome b reductase fragment of nitrate reductase. J Biol Chem 270: 24067–24072

    Article  CAS  PubMed  Google Scholar 

  • Ratnam K, Shiraishi N, Campbell WH and Hille R (1997) Spectroscopic and kinetic characterization of the recombinant cytochrome c reductase fragment of nitrate reductase: identification of the rate limiting catalytic step. J Biol Chem 272: 2122–2128

    CAS  PubMed  Google Scholar 

  • Redinbaugh MG and Campbell WH (1991) Higher plant responses to environmental nitrate. Physiol Plant 82: 640–650

    Article  CAS  Google Scholar 

  • Redinbaugh MG and Campbell WH (1993) Glutamine synthetase and ferredoxin-dependent glutamate synthase expression in the maize (Zea mays) root primary response to nitrate. Plant Physiol 101: 1249–1255

    CAS  PubMed  Google Scholar 

  • Redinbaugh MG and Campbell WH (1998) Nitrate regulation of the oxidative pentose phosphate pathway in maize root plastids: Induction of 6-phosphogluconate activity, protein and transcript levels. Plant Sci 134: 129–140

    Article  CAS  Google Scholar 

  • Remmler JL and Campbell WH (1986) Regulation of corn leaf nitrate reductase: II. Synthesis and turnover of the enzyme’s activity and protein. Plant Physiol 80: 442–447

    CAS  Google Scholar 

  • Ritchie SW, Redinbaugh MG, Shiraishi N, Verba JM and Campbell WH (1994) Identification of a maize root transcript expressed in the primary response to nitrate: Characterization of a cDNA with homology to ferredoxin-NADP+ reductase. Plant Mol Biol 26: 679–690

    Article  CAS  PubMed  Google Scholar 

  • Sakakibara H, Kobayashi K, Deji A and Sugiyama T (1997) Partial characterization of the signaling pathway for the nitrate-dependent expression of genes for nitrogen-assimilatory enzymes using detached leaves of maize. Plant Cell Physiol 38: 837–843

    CAS  Google Scholar 

  • Scheible W-R, Gonzales-Fontes A, Lauerer M, Muller-Rober B, Caboche M and Stitt M (1997) Nitrate acts as a signal to induce organic acid metabolism and repress starch metabolism in tobacco. Plant Cell 9: 783–798

    Article  CAS  PubMed  Google Scholar 

  • Schwarz G, Boxer DH and Mendel RR (1997) Molybdenum cofactor biosynthesis. The plant protein Cnxl binds molybdopterin with high affinity. J Biol Chem 272: 26811–26814

    CAS  PubMed  Google Scholar 

  • Smil V (1997) Global Population and the Nitrogen Cycle. Sci Amer 277: 76–81

    CAS  Google Scholar 

  • Stitt M (1999) Nitrate regulation of metabolism and growth. Curr Opin Plant Biol 2: 178–186

    Article  CAS  PubMed  Google Scholar 

  • Su W, Huber SC and Crawford NM (1996) Identification in vitro of a post-translational regulatory site in the hinge 1 region of Arabidopsis nitrate reductase. Plant Cell 8: 519–527

    Article  CAS  PubMed  Google Scholar 

  • Su W, Mertens JA, Kanamaru K, Campbell WH and Crawford NM (1997) Analysis of wild-type and mutant plant nitrate reductase expressed in the methylotrophic yeast Pichia pastoris. Plant Physiol 115: 1135–1143

    Article  CAS  PubMed  Google Scholar 

  • Sugden C, Donaghy PG, Halford NG and Hardie DG (1999) Two SNF1-related protein kinases from spinach leaf phosphorylate and inactivate 3-hydroxy-3-methylglutaryl-coenzyme A reductase, nitrate reductase, and sucrose phosphate synthase in vitro. Plant Physiol 120: 257–274

    Article  CAS  PubMed  Google Scholar 

  • Toroser D, Plaut Z and Huber SC (2000) Regulation of a plant SNF1-related protein kinase by glucose-6-phosphate. Plant Physiol 123: 403–412

    Article  CAS  PubMed  Google Scholar 

  • Vidmar JJ, Zhuo D, Siddiqi MY, Schjoerring JK, Touraine B and Glass AMD (2000) Regulation of high-affinity nitrate transporter genes and high-affinity nitrate influx by nitrogen pools in roots of barley. Plant Physiol 123: 307–318

    Article  CAS  PubMed  Google Scholar 

  • Vincentz M and Caboche M (1991) Constitutive expression of nitrate reductase allows for normal growth and development of Nicotiana plumbaginifolia plants. EMBO J 10: 1027–1035

    CAS  PubMed  Google Scholar 

  • Vitousek PM, Mooney HA, Lubchenco J and Melillo JM (1997) Human Domination of Earth’s Ecosystems. Science 277: 494–499

    Article  CAS  Google Scholar 

  • Wang R, Guegler K, LaBrie ST and Crawford NM (2000) Genomic analysis of a nutrient response in Arabidopsis reveals diverse expression patterns and novel metabolic and potential regulatory genes induced by nitrate. Plant Cell 12: 1491–1509

    CAS  PubMed  Google Scholar 

  • Weiner H and Kaiser WM (1999) 14-3-3 proteins control proteolysis of nitrate reductase in spinach leaves. FEBS Lett 455: 75–78

    Article  CAS  PubMed  Google Scholar 

  • Yamasaki H and Sakihama Y (2000) Simultaneous production of nitric oxide and peroxynitrite by plant nitrate reductase: in vitro evidence for the NR-dependent formation of active nitrogen species. FEBS Lett 468: 89–92

    Article  CAS  PubMed  Google Scholar 

  • Zhang H and Forde BG (1998) An Arabidopsis MADS box gene that controls nutrient-induced changes in root architecture. Science 279: 407–409

    Article  CAS  PubMed  Google Scholar 

  • Zhang H, Forde BG (2000) Regulation of Arabidopsis root development by nitrate availability. J Exp Bot 51: 51–59

    Article  CAS  PubMed  Google Scholar 

  • Zhang H, Jennings A, Barlow PW and Forde BG (1999) Dual pathways for regulation of root branching by nitrate. Proc Natl Acad Sci USA 96: 6529–6534

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biological Sciences, Michigan Technological University, Houghton, MI, 49931-1295, USA

    Wilbur H. Campbell

Authors
  1. Wilbur H. Campbell
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Crop Performance and Improvement Division, IACR-Rothamsted, Harpenden, UK

    Christine H. Foyer

  2. Institut de la Biotechnologie des Plantes, Université Denis Diderot Paris VII, Orsay, France

    Graham Noctor

Rights and permissions

Reprints and permissions

Copyright information

© 2002 Kluwer Academic Publishers

About this chapter

Cite this chapter

Campbell, W.H. (2002). Molecular Control of Nitrate Reductase and Other Enzymes Involved in Nitrate Assimilation. In: Foyer, C.H., Noctor, G. (eds) Photosynthetic Nitrogen Assimilation and Associated Carbon and Respiratory Metabolism. Advances in Photosynthesis and Respiration, vol 12. Springer, Dordrecht. https://doi.org/10.1007/0-306-48138-3_3

Download citation

  • DOI: https://doi.org/10.1007/0-306-48138-3_3

  • Publisher Name: Springer, Dordrecht

  • Print ISBN: 978-0-7923-6336-1

  • Online ISBN: 978-0-306-48138-3

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation