Abstract
Previous studies have shown that expression of the gdhA gene, encoding NADP-linked glutamate dehydrogenase (NADP-GDH), in Aspergillus nidulans is regulated by the major nitrogen regulatory protein AreA and its co-activator TamA. We show here that loss of TamA function has a more severe effect on the levels of gdhA expression than loss of AreA function. Using TamA as the bait in a yeast two-hybrid screen, we have identified a second protein that interacts with TamA. Sequencing analysis and functional studies have shown that this protein, designated LeuB, is a transcriptional activator with similar function to the homologous Leu3p of Saccharomyces cerevisiae. Inactivation of leuB revealed that this gene is involved in the regulation of gdhA, and an areA; leuB double mutant was shown to have similar NADP-GDH levels to a tamA single mutant. The requirement for TamA function to promote gdhA expression is likely to be due to its dual interaction with AreA and LeuB.
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Acknowledgements
We acknowledge the support of the Australian Research Council and the awards of a University of Melbourne Career Interruption Fellowship to R.P. and a Melbourne Research Scholarship to B.J.M. We thank Nicole Kennon for assistance with NADP-GDH assays, and Sarah Pitt and Jairus Bowne for technical assistance in cloning the luA gene
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Communicated by P. J. Punt
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Polotnianka, R., Monahan, B.J., Hynes, M.J. et al. TamA interacts with LeuB, the homologue of Saccharomyces cerevisiae Leu3p, to regulate gdhA expression in Aspergillus nidulans. Mol Genet Genomics 272, 452–459 (2004). https://doi.org/10.1007/s00438-004-1073-x
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DOI: https://doi.org/10.1007/s00438-004-1073-x