title: Regulation of the NADP-glutamate dehydrogenase gene gdhA in Aspergillus nidulans by the Zn(II)2Cys6 transcription factor LeuB.
abstract: NADP-dependent glutamate dehydrogenase (NADP-GDH) is a key enzyme in the assimilation of alternative nitrogen nutrient sources through ammonium in fungi. In Aspergillus nidulans, NADP-GDH is encoded by gdhA. Several transcription factors are known to regulate gdhA expression, including AreA, the major transcription activator of nitrogen metabolic genes, and TamA, a co-activator of AreA. TamA also interacts with LeuB, the regulator of leucine biosynthesis. We have investigated the effects of leucine biosynthesis on gdhA regulation, and found that leucine regulates the levels of NADP-GDH activity and gdhA expression. We show, using mutants with perturbed levels of α-isopropylmalate (α-IPM), that this leucine biosynthesis intermediate affects gdhA regulation. Leucine regulation of gdhA requires a functional LeuB with an intact Zn(II)2Cys6 DNA-binding domain. By analysing the prevalence of putative LeuB DNA-binding sites in promoters of gdhA orthologues we predict broad conservation of leucine regulation of NADP-GDH expression within ascomycetes except in the fusaria and fission yeasts. Using promoter mutations in gdhA-lacZ reporter genes we identified two sites of action for LeuB within the A. nidulans gdhA promoter. These two sites lack sequence identity, with one site conforming to the predicted LeuB DNA-binding site consensus motif, whereas the second site is a novel regulatory sequence element conserved in Aspergillus gdhA promoters. These data suggest that LeuB regulates NADP-GDH expression in response to leucine levels, which may act as an important sensor of nitrogen availability.
authors.affiliation: Department of Genetics, University of Melbourne, Parkville, VIC 3010, Australia.|Department of Plant Pathology, Kansas State University, 4024 Throckmorton Plant Sciences Center, Manhattan, KS 66506, USA.
authors.forename: Damien J
authors.initials: DJ
authors.lastname: Downes
authors.affiliation: Department of Genetics, University of Melbourne, Parkville, VIC 3010, Australia.
authors.forename: Meryl A
authors.initials: MA
authors.lastname: Davis
authors.affiliation: Department of Genetics, University of Melbourne, Parkville, VIC 3010, Australia.
authors.forename: Sara D
authors.initials: SD
authors.lastname: Kreutzberger
authors.affiliation: Department of Genetics, University of Melbourne, Parkville, VIC 3010, Australia.
authors.forename: Brendan L
authors.initials: BL
authors.lastname: Taig
authors.affiliation: Department of Genetics, University of Melbourne, Parkville, VIC 3010, Australia.|Department of Plant Pathology, Kansas State University, 4024 Throckmorton Plant Sciences Center, Manhattan, KS 66506, USA.
authors.forename: Richard B
authors.initials: RB
authors.lastname: Todd
mesh_terms: D019677:Artificial Gene Fusion; D001233:Aspergillus nidulans / Q000201:enzymology*; D001665:Binding Sites / Q000235:genetics; D004252:DNA Mutational Analysis; D015966:Gene Expression Regulation, Fungal; D017930:Genes, Reporter; D018086:Glutamate Dehydrogenase (NADP+) / Q000096:biosynthesis*; D007930:Leucine / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
chemical_list: D014157:Transcription Factors; D018086:Glutamate Dehydrogenase (NADP+); D001616:beta-Galactosidase; D007930:Leucine