PMID: 8606156Apr 1, 1996Paper

The CCAAT box-binding factor stimulates ammonium assimilation in Saccharomyces cerevisiae, defining a new cross-pathway regulation between nitrogen and carbon metabolisms

Journal of Bacteriology
V D DangB Daignan-Fornier

Abstract

In Saccharomyces cerevisiae, carbon and nitrogen metabolisms are connected via the incorporation of ammonia into glutamate; this reaction is catalyzed by the NADP-dependent glutamate dehydrogenase (NADP-GDH) encoded by the GDH1 gene. In this report, we show that the GDH1 gene requires the CCAAT box-binding activator (HAP complex) for optimal expression. This conclusion is based on several lines of evidence: (1) overexpression of GDH1 can correct the growth defect of hap2 and hap3 mutants on ammonium sulfate as a nitrogen source, (ii) Northern (RNA) blot analysis shows that the steady-state level of GDH1 mRNA is strongly lowered in a hap2 mutant, (iii) expression of a GDH1-lacZ fusion is drastically reduced in hap mutants, (iv) NADP-GDH activity is several times lower in the hap mutants compared with that in the isogenic wild-type strain, and finally, (v) site-directed mutagenesis of two consensual HAP binding sites in the GDH1 promoter strongly reduces expression of GDH1 and makes it HAP independent. Expression of GDH1 is also regulated by the carbon source, i.e., expression is higher on lactate than on ethanol, glycerol, or galactose, with the lowest expression being found on glucose. Finally, we show that a hap2 mutation does...Continue Reading

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Citations

Dec 16, 1997·Proceedings of the National Academy of Sciences of the United States of America·D A LashkariR W Davis
Oct 14, 2003·Nature Biotechnology·Ziv Bar-JosephDavid K Gifford
Feb 10, 2004·Molecular Biology of the Cell·Petra de GraafPaul M P van Bergen en Henegouwen
Mar 23, 2005·FEMS Yeast Research·Edna Maria Morais OliveiraElba Pinto da Silva Bon
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Aug 1, 2008·Expert Opinion on Drug Discovery·Anton Yuryev
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Jun 11, 2002·Biochemical and Biophysical Research Communications·Lina RiegoAlicia González
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