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Ammonia assimilation in Saccharomyces cerevisiae as mediated by the two glutamate dehydrogenases

Evidence for the gdhA locus being a structural gene for the NADP-dependent glutamate dehydrogenase

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Summary

Mutants lacking NADP-linked glutamate dehydrogenase (NADP-GDH) activity have been isolated by several procedures.

Complementation tests in diploids as well as tetrad analysis show that they map within a short chromosome segment, the gdhA locus, which is allelic to the ure1 locus described previously.

That the gdhA locus is a structural gene for NADP-GDH is supported by two kinds of evidence. First, intracistronic complementation, as well as negative complementation were observed between some of the gdhA- mutants. This is in agreement with the multimeric structure of the NADP-GDH in Saccharomyces cerevisiae shown by Venard and Fourcade (1972). Secondly, some of the mutants at the gdhA locus have a NADP-GDH with modified properties, including: five-fold higher Km for 2-oxoglutarate, hundred-fold higher Km for NH +4 , loss of inhibition by excess of substrate (2-oxoglutarate), and lower thermostability.

Mutants with derepressed NAD-GDH activity have been isolated from gdhA- strains on the basis of their faster growth on ammonia as sole nitrogen source. They define the gdhCR locus, which is allelic to ure2 and usu described previously. This is a strong indication that residual growth of the gdhA- mutants on ammonia as sole nitrogen source is due to the NAD-GDH activity.

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Author notes

  1. Malgorzata Piotrowska (Holder of an EMBO fellowship)

    Present address: Dept. of Genetics, Institute of Biochemistry and Biophysics, Rakowiecka 36, Warszawa, Poland

Authors and Affiliations

  1. Laboratoire de Microbiologie, Faculté des Sciences, Université Libre de Bruxelles, Bruxelles, Belgique

    Marcelle Grenson, Evelyne Dubois & Malgorzata Piotrowska (Holder of an EMBO fellowship)

  2. Institut de Recherches du C.E.R.I.A., Bruxelles, Belgique

    Marcelle Grenson, Evelyne Dubois & Malgorzata Piotrowska (Holder of an EMBO fellowship)

  3. Laboratoire de Génétique physiologique, Institut de Biologie moléculaire et cellulaire, Strasbourg, France

    Robert Drillien & Michel Aigle

Authors
  1. Marcelle Grenson
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  2. Evelyne Dubois
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  3. Malgorzata Piotrowska
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  4. Robert Drillien
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  5. Michel Aigle
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Additional information

Communicated by W. Gajewski

Aspirant du Fonds National de la Recherche Scientifique Belge.

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Grenson, M., Dubois, E., Piotrowska, M. et al. Ammonia assimilation in Saccharomyces cerevisiae as mediated by the two glutamate dehydrogenases. Molec. Gen. Genet. 128, 73–85 (1974). https://doi.org/10.1007/BF00267295

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  • DOI: https://doi.org/10.1007/BF00267295

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