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A conditional-lethal molybdopterin-defective mutant of barley

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Summary

The molybdenum cofactor of the barley mutant R9401 is not able to reconstitute NADPH nitrate reductase activity from extracts of the N. crassa nit-1 mutant nor is it able to effect dimerisation of the nitrate reductase subunits present in the R9401 mutant. Unphysiologically high levels of molybdate cannot restore nitrate reductase and xanthine dehydrogenase activity to mutant R9401 in vivo nor reactivate the Mo-co factor in vitro. The results indicate that the defect in mutant R9401 lies in the pathway leading to the formation of a functional molybdopterin moiety and that the same nuclear gene is involved in the synthesis of both shoot and root molybdenum cofactor.

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Abbreviations

BSA:

bovine serum albumen

GSH:

glutathione (reduced)

NEM :

N-ethylmaleimide

References

  • Bright SWJ, Norbury PB, Franklin J, Kirk DW, Wray JL (1983) A conditional-lethal cnx-type nitrate reductase-deficient barley mutant. Mol Gen Genet 189:240–244

    Google Scholar 

  • Brown J, Small IS, Wray JL (1981) Age-dependent conversion of nitrate reductase to cytochrome c reductase species in barley leaf extracts. Phytochem 20:389–398

    Google Scholar 

  • Buchanan RJ, Wray JL (1982) Isolation of molybdenum cofactor defective cell lines of Nicotiana tabacum. Mol Gen Genet 188:228–234

    Google Scholar 

  • Campbell JMcA, Wray JL (1983) Purification of barley nitrate reductase and demonstration of nicked subunits Phytochem 22:2375–2382

    Google Scholar 

  • Cove DJ (1979) Genetic studies of nitrate assimilation in Aspergillus nidulans. Biol Rev 54:291–327

    Google Scholar 

  • Dunn-Coleman NS, Smarrelli J, Garrett RH (1984) Nitrate assimilation in eukaryotic cells. Int Rev Cytol 92:1–50

    Google Scholar 

  • Ferrario S, Hirel B, Gadal P (1983) Immunological characterisation of nitrate reductase from spinach leaves and roots. Biochem Biophys Res Commun 113:733–737

    Google Scholar 

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

    Google Scholar 

  • Gewitz H-S, Piefke J, Vennesland B (1981) Purification and characterisation of demolybdo nitrate reductase (NADH cytochrome c oxidoreductase) of Chlorella vulgaris. J. Biol Chem 256:11527–11531

    Google Scholar 

  • Hewitt EJ, Notton BA, Rucklidge GJ (1977) Formation of nitrate reductase by recombination of apoprotein fraction from molybdenum deficient plants with a molybdenum containing complex J Less Common Metals 54:537–545

    Google Scholar 

  • Johnson JL (1980) The molybdenum cofactor common to nitrate reductase, xanthine dehydrogenase and sulfite oxidase. In: Coughlan M (ed) Molybdenum and molybdenum-containing enzymes. Pergamon Press, Oxford, p 345–385

    Google Scholar 

  • Johnson JL, Rajagopalan KV (1982) Structural and metabolic relationship between the molybdenum cofactor and urothione. Proc Natl Acad Sci USA 79:6856–6860

    Google Scholar 

  • Johnson JL, Hainline BE, Rajagopalan KV (1980) Characterisation of the molybdenum cofactor of sulphite oxidase, xanthine oxidase and nitrate reductase. Identification of a pteridine as a structural component J Biol Chem 255:1783–1786

    Google Scholar 

  • Kleinhofs A, Taylor J, Kuo TM, Somers DA, Warner RL (1983) Nitrate reductase genes as selectable markers for plant cell transformation. In: Lurquin PF, Kleinhofs A (eds) Genetic engineering in eukaryotes. Plenum Publishing Coroporation, New York, p 215–231

    Google Scholar 

  • Kuo T, Kleinhofs A, Warner RL (1980) Purification and partial characterisation of nitrate reductase from barley leaves. Plant Sci Lett 17:371–381

    Google Scholar 

  • Marton L, Sidorov V, Biasini G, Maliga P (1982) Complementation in somatic hybrids indicates four types of nitrate reductase deficient lines in Nicotiana plumbaginifolia. Mol Gen Genet 187:1–3

    Google Scholar 

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

    Google Scholar 

  • Mendel RR, Müller AJ (1985) Repair in vitro of nitrate reductasedeficient tobacco mutants (cnxA) by molybdate and by molybdenum cofactor. Planta 163:370–375

    Google Scholar 

  • Mendel RR, Alikulov ZA, Lvov NP, Müller AJ (1981) Presence of the molybdenum cofactor in nitrate reductase-deficient mutant cell lines of Nicotiana tabacum. Mol Gen Genet 181:395–399

    Google Scholar 

  • Mendel RR, Marton L, Wray JL (1982) Biochemical evidence for three types of molybdenum cofactor-defective mutants (cnx) in Nicotiana. Proc Int Symp on “Nitrate Assimilation — Molecular and Genetic Aspects”, Gatersleben, GDR, P 16

  • Mendel RR, Buchanan RJ, Wray JL (1984) Characterisation of a new type of molybdenum cofactor-mutant in cell cultures of Nicotiana tabacum. Mol Gen Genet 195:186–189

    Google Scholar 

  • Mendel RR, Kirk DW, Wray JL (1985) Assay of molybdenum — cofactor of barley. Phytochemistry 24:1631–1634

    Google Scholar 

  • Müller AJ, Grafe R (1978) Isolation and characterisation of cell lines of Nicotiana tabacum lacking nitrate reductase. Mol Gen Genet 161:67–76

    Google Scholar 

  • Narayanan KR, Somers DA, Kleinhofs A, Warner RL (1983) Nature of cytochrome c reductase in nitrate reductase-deficient mutants in barley. Mol Gen Genet 190:222–226

    Google Scholar 

  • Nason A, Antoine A-D, Ketchum PA, Frazier III WA, Lee DK (1970) Formation of assimilatory nitrate reductase by in vitro intercistronic complementation in Neurospora crassa. Proc Natl Acad Sci USA 65:137–144

    Google Scholar 

  • Nason A, Lee KY, Ketchum PA, Lamberti A, DeVries J (1971) In vitro formation of assimilatory reduced nicotinamide dinucleotide phosphate nitrate reductase from a Neurospora mutant and a component of molybdenum-enzymes. Proc Natl Acad Sci USA 68:3242–3246

    Google Scholar 

  • Ramadoss CS, Shen TC, Vennesland B (1981) Molybdenum insertion in vitro in demolybdo nitrate reductase of Chlorella vulgaris. J Biol Chem 256:11532–11537

    Google Scholar 

  • Redinbaugh MG, Campbell WH (1985) Quaternary structure and composition of squash NADH: nitrate reductase. J Biol Chem 260:3380–3385

    Google Scholar 

  • Small IS, Wray JL (1980) NADH nitrate reductase and related NADH cytochrome c reductase species in barley. Phytochemistry 19:387–394

    Google Scholar 

  • Somers DA, Kuo TM, Kleinhofs A, Warner RL (1983) Nitrate reductase-deficient mutants in barley. Immunoelectrophoretic characterisation. Plant Physiol 71:145–149

    Google Scholar 

  • Tokarev EI, Shumny VK (1977) Clarification of barley mutants with lowered nitrate reductase activity after treatment of the grain with ethylmethanesulphonate. Genetika (Moscow) 13:2097–2103

    Google Scholar 

  • Wahl RC, Hageman RV, Rajagopalan KV (1984) The relationship of Mo, molybdopterin, and the cyanolyzable sulfur in the Mo cofactor. Arch Biochem Biophys 230:264–273

    Google Scholar 

  • Warner RL, Lin CJ, Kleinhofs A (1977) Nitrate reductase deficient mutants in barley. Nature 269:406–407

    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 

  • Xuan LT, Grafe R, MÜller AJ (1983) Complementation of nitrate reductase deficient mutants in somatic hybrids between Nicotiana species. In: Potrykus I (eds), Protoplasts 1983 (Poster Proceedings) Birkhauser, Basel, pp 76–77

    Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry and Microbiology, University of St, Andrews, North Street, KY16 9AL, St. Andrews, Fife

    J. L. Wray, B. Steven & D. W. Kirk

  2. Department of Biochemistry, Rothamsted Experimental Station, AL5 2JQ, Harpenden, Hertfordshire, United Kingdom

    B. Steven & S. W. J. Bright

Authors
  1. J. L. Wray
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  2. B. Steven
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  3. D. W. Kirk
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  4. S. W. J. Bright
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Communicated by G. Melchers

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Wray, J.L., Steven, B., Kirk, D.W. et al. A conditional-lethal molybdopterin-defective mutant of barley. Molec Gen Genet 201, 462–466 (1985). https://doi.org/10.1007/BF00331340

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  • DOI: https://doi.org/10.1007/BF00331340

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