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Biochemical characterization of the molybdenum cofactor mutants of Neurospora crassa: in vivo and in vitro reconstitution of NADPH-nitrate reductase activity

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Summary

Molybdenum cofactor (MoCo) mutants of Neurospora crassa lack both NADPH-nitrate reductase and xanthine dehydrogenase activity. In vivo and in vitro studies to further characterize these mutants are now reported. The MoCo mutants nit-9A and nit-9B are capable of growing, albeit poorly, with nitrate as the sole nitrogen source, provided high levels of molybdate are present. The MoCo mutants nit-9A, nit-9B and nit-9C, but not nit-1, nit-7 or nit-8, have significant levels of NADPH-nitrate reductase when grown in nitrate medium containing 30 mM molybdate. In vitro reconstitution experiments using cell free extracts of the N. crassa MoCo mutants and E. coli HB101 as a source of wild-type MoCo were performed. MoCo from E. coli was capable of reconstituting NADPH-nitrate reductase activity to nit-1, nit-7 and nit-8. Molybdate is required for the in vitro reconstitution of NADPH-nitrate reductase activity. It was not possible to in vitro reconstitute NADPH-nitrate reductase activity in the MoCo mutants nit-9A, nit-9B or nit-9C.

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  1. Nigel Stuart Dunn-Coleman (Visiting Scientist)

    Present address: Central Research and Development Department, Du Pont Experimental Station, 19898, Wilmington, Delaware, USA

Authors and Affiliations

  1. International Plant Research Institute, 94070, San Carlos, California, USA

    Nigel Stuart Dunn-Coleman (Visiting Scientist)

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  1. Nigel Stuart Dunn-Coleman
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Dunn-Coleman, N.S. Biochemical characterization of the molybdenum cofactor mutants of Neurospora crassa: in vivo and in vitro reconstitution of NADPH-nitrate reductase activity. Curr Genet 8, 581–588 (1984). https://doi.org/10.1007/BF00395703

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  • DOI: https://doi.org/10.1007/BF00395703

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