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Biochemical studies on the Nit mutants of Neurospora crassa

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Summary

One allele at each of the five nit loci in Neurospora crassa together with the wild type strain have been compared on various nitrogen sources with regard to (i) their growth characteristics (ii) the level of nitrate reductase and its associated activities (reduced benzyl viologen nitrate reductase and cytochrome c reductase) (iii) the level of nitrite reductase and (iv) their ability to take up nitrite from the surrounding medium. Results are consistent with the hypothesis that nit-3 is the structural gene for nitrate reductase, nit-1 specifies in part a molybdenum containing moiety which is responsible for the nit-3 gene product dimerising to form nitrate reductase, nit-4 and nit-5 are regulator genes whose products are involved in the induction of both nitrate reductase and nitrite reductase and nit-2 codes for a generalised ammonium activated repressor protein. Studies on the induction of nitrate reductase (and its associated activities) and nitrite reductase in wild type, nit-1 and nit-3 in the presence of either nitrate or nitrite suggest that each enzyme may be regulated independently of the other and that nitrite could be true co-inducer of the assimilatory pathway. Nitrite uptake experiments with nit-2, nit-4 and nit-5 strains show that whereas nit-4 and nit-5 are freely permeable to this molecule, it is unable to enter the nit-2 mycelium.

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References

  • Ackers, G.: A new calibration procedure for gel filtration columns. J. biol. Chem. 242, 3237–3238 (1967)

    Google Scholar 

  • Antoine, A.D.: Furification and properties of the nitrate reductase isolated from Neurospora crassa mutant nit-3. Kinetics, molecular weight determination and cytochrome involvement. Biochemistry 13, 2289–2294 (1974)

    Google Scholar 

  • Arst, H.N., Cove, D.J.: Nitrogen metabolite repression in Aspergillus nidulans. Molec. gen. Genet. 126, 111–141 (1973)

    Google Scholar 

  • Arst, H.N., MacDonald, D.W., Cove D.J.: Molybdate metabolism in Aspergillus nidulans. I. Mutations affecting nitrate reductase and/or xanthine dehydrogenase. Molec. gen. Genet. 108, 129–145 (1970)

    Google Scholar 

  • Beers, R.F., Sizer, I.W.: A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J. biol. Chem. 195, 133–140 (1952)

    Google Scholar 

  • Cove, D.J.: The induction and repression of nitrate reductase in the fungus Aspergillus nidulans. Biochim. biophys. Acta (Amst.) 113, 51–56 (1966)

    Google Scholar 

  • Cove, D.J.: Control of gene action in Aspergillus nidulans. Proc. roy. Soc. B 176, 267–275 (1970)

    Google Scholar 

  • Cove, D.J., Coddington, A.: Purifiction of nitrate reductase and cytochrome c reductase from Aspergillus nidulans. Biochim. biophys. Acta (Amst.) 110, 312–318 (1965)

    Google Scholar 

  • Cove, D.J., Pateman, J.A.: Autoregulation of the synthesis of nitrate reductase in Aspergillus nidulans. J. Bact 97, 1374–1378 (1969)

    Google Scholar 

  • Garrett, R. H.: The induction of nitrite reductase in Neurospora crassa. Biochim. biophys. Acta (Amst.) 264, 481–489 (1972)

    Google Scholar 

  • Garrett, R.H., Nason, A.: Further purification and properties of Neurospora nitrate reductase. J. biol. Chem. 244, 2870–2882 (1969)

    Google Scholar 

  • Kaplan, D., Roth-Bejeramo, N., Lips, S.H.: Nitrate reductase as a product inducible enzyme. Europ. J. Biochem. 49, 393–398 (1974)

    Google Scholar 

  • Ketchum, P.A., Cambier, H.Y., Frazier, W.A., Madansky, C.H., Nason, A.: In vitro assembly of Neurospora assimilatory nitrate reductase from protein sub-units of a Neurospora mutant and the xanthine oxidising or aldehyde oxidase systems of higher animaly. Proc. nat. Acad. Sci. (Wash.) 66, 1016–1023 (1970)

    Google Scholar 

  • Kinsky, S.C., McElroy, W.D.: Neurospora, nitrate reductase: The role of phosphate flavine and cytochrome c reductase. Arch. Biochem. Biophys. 73, 466–483 (1958)

    Google Scholar 

  • Lee, K.-Y., Pan, S.-S., Erickson, R., Nason, A.: Involvement of molybdenum and iron in the in vitro assembly of assimilatory nitrate reductase utilizing Neurospora mutant nit-1. J. biol. Chem. 249, 3941–3952 (1974)

    Google Scholar 

  • Lowry, O.H., Roseborough, N.J., Farr, A.L., Randall, R.J.: Protein measurement with the Folin phenol reagent. J. biol. Chem. 193, 265–275 (1951)

    Google Scholar 

  • MacDonald, D.W., Coddington, A.: Properties of the assimilatory nitrate reductase from Asperigillus nidulans. Europ. J. Biochem. 46, 169–178 (1974)

    Google Scholar 

  • MacDonald, D.W., Cove, D.J., Coddington, A.: Cytochrome c reductases from wild type and mutant strains of Aspergillus nidulans. Molec. gen. Genet. 128, 187–199 (1974)

    Google Scholar 

  • Martin, R.G., Ames, B.N.: A method for determining the sedimentation behaviour of enzymes. Application to protein mixtures. J. biol. Chem. 236, 1372–1379 (1961)

    Google Scholar 

  • Nason, A., Antoine, A.D., Ketchum, P.A. Frazier, W.A., Lee, D.K.: Formation of an assimilatory nitrate reductase by in vitro intercistronic complementation in Neurospora crassa. Proc. nat. Acad. Sci. (Wash.) 65, 133–144 (1970)

    Google Scholar 

  • Nason, A., Evans, H.J.: Triphospho-pyridine nucleotide nitrate reductase in Neurospora. J. biol. Chem. 202, 655–673 (1953)

    Google Scholar 

  • Nason, A., Lee, K-Y, Pan, S-S, Ketchum, P.A., Lambert, A. DeVries, J.: in vitro formation of assimilatory reduced nicotinamide adenine dinucleotide phosphate nitrate reductase from a Neurospora mutant and a component of molybdenum enzymes. Proc. nat. Acad. Sci. (Wash.) 68, 3242–3246 (1971)

    Google Scholar 

  • Nicholas, D.J.D., Nason, A.: Mechanism of action of nitrate reductase from Neurospora. J. biol. Chem. 211, 183–197 (1954)

    Google Scholar 

  • Pateman, J.A., Rever, B.M., Cove, D.J.: Genetical and biochemical studies of nitrate reduction in Aspergillus nidulans. Biochem. J. 104, 103–111 (1967)

    Google Scholar 

  • Scarborough, G.A.: Sugar transport in Neurospora crassa. J. biol. Chem. 245, 1694–1698 (1970)

    Google Scholar 

  • Schloemer, R.H., Garrett, R.H.: Nitrate transport system in Neurospora crassa. J. Bact. 118, 259–269 (1974a)

    Google Scholar 

  • Schloemer, R.H., Garrett, R.H.: Uptake of nitrite by Neurospora crassa. J. Bact. 118, 270–274 (1974b)

    Google Scholar 

  • Siegel, L.M., Monty, K.J.: Determination of molecular weights and frictional ratios of proteins in impure systems by the use of gel filtration and density gradient centrifugation: Applications to crude preparations of sulfite and hydroxylamine reductases. Biochim. biophys. Acta (Amst.) 112, 346–362 (1966)

    Google Scholar 

  • Sorger, G.J.: Simultaneous induction and repression of nitrate reductase and TPNH-cytochrome c reductase in Neurospora crassa. Biochim. biophys. Acta (Amst.) 99, 234–245 (1965)

    Google Scholar 

  • Sorger, G.J.: Nitrate reductase electron transport systems in mutant and in wild type strains of Neurospora. Biochim. biophys. Acta (Amst.) 118, 484–494 (1966)

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Sorger, G.J., Giles, N.H.: Genetic control of nitrate reductase in Neurospora crassa. Genetics 52, 777–788 (1965)

    Google Scholar 

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

    Google Scholar 

  • Vallee, B.L., Hoch, F.L.: Zinc, a component of yeast alcohol dehydrogenase. Proc. nat. Acad. Sci. (Wash.) 41, 327–338 (1955)

    Google Scholar 

  • Vogel, H.J., Bonner, D.M.: Microbial Genetics Bull. 13, 42 (1956)

    Google Scholar 

  • Wiley, W.R., Matchett, W.H.: Tryptophan transport in Neurospora crassa. I. Specificity and kinetics. J. Bact. 92, 1698–1705 (1966)

    Google Scholar 

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  1. School of Biological Sciences, University of East Anglia, NR4 7TJ, Norwich, Great Britain

    Alan Coddington

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  1. Alan Coddington
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Communicated by W. Gajewski

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Coddington, A. Biochemical studies on the Nit mutants of Neurospora crassa . Molec. Gen. Genet. 145, 195–206 (1976). https://doi.org/10.1007/BF00269594

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  • DOI: https://doi.org/10.1007/BF00269594

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