Abstract
InAspergillus nidulans the positive-acting, wide domain regulatory geneareA mediates nitrogen metabolite repression. Previous analysis demonstrated that the C-terminal 153 residues of theareA product (AREA) are inessential for at least partial expression of most genes subject to regulation byareA. Paradoxically,areA r 2, a −1 frameshift replacing the wild-type 122 C-terminal residues with a mutant peptide of 117 amino acids, leads to general loss of function. To determine the basis for theareA r 2 mutant phenotype, and as a means of delineating functional domains within the C-terminal region of AREA, we have selected and characterisedareA r 2 revertants. Deletion analysis, utilising direct gene replacement, extended this analysis. A mutantareA product truncated immediately after the last residue of the highly conserved GATA (DNA-binding) domain retains partial function. TheareA r 2 product retains some function with respect to the expression ofuaZ (encoding urate oxidase) and the mutant allele is partially dominant with respect to nitrate reductase levels. Consistent with theareA r 2 product having a debilitating biological activity, we have demonstrated that a polypeptide containing both the wild-type DNA-binding domain and the mutant C-terminus of AREA2 is able to bind DNA in vitro but no longer shows specificity for GATA sequences.
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Communicated by C. A. M. J. J. van den Hondel
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Platt, A., Ravagnani, A., Arst, H. et al. Mutational analysis of the C-terminal region of AREA, the transcription factor mediating nitrogen metabolite repression inAspergillus nidulans . Molec. Gen. Genet. 250, 106–114 (1996). https://doi.org/10.1007/BF02191830
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DOI: https://doi.org/10.1007/BF02191830