Abstract
The major nitrogen regulatory gene, areA, was cloned from Aspergillus parasiticus. It encoded a polypeptide of 864 amino acids which contained a nuclear localization signal (NLS), a highly acidic region from positions 497 to 542, a Cys–X 2–Cys–X 17–Cys–X 2–Cys DNA-binding motif and a conserved carboxy-terminus. Electrophoretic mobility shift assays suggested that the A. parasiticus AREA DNA-binding domain fusion protein bound cooperatively to single GATA elements in the A. parasiticus niaD– niiA intergenic region. AREA also bound to the aflR– aflJ intergenic region of the aflatoxin biosynthesis gene cluster. Regions of areA were fused to a yeast GAL4 DNA-binding domain coding region to localize putative transcription activation domain(s) of AREA based on activation of the GAL1(p)::lacZ reporter gene expression. The portion between NLS and the acidic domain demonstrated 16–20-fold higher activation activities than other portions of AREA, which suggests that the transcription activation domain is located in this region.
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Schedule a callMore From: Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression
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