Characterization of the Aspergillus parasiticus major nitrogen regulatory gene, areA.
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.
Chang
P K
PK
Southern Regional Research Center, Agricultural Research Service, US Department of Agriculture, 1100 Robert E. Lee Boulevard, New Orleans, LA, USA. pkchang@nola.srrc.usda.gov
Yu
J
J
Bhatnagar
D
D
Cleveland
T E
TE
D001230:Aspergillus / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D009566:Nitrates / Q000378:metabolism; D014157:Transcription Factors / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D014158:Transcription, Genetic
C079728:AreA protein, Aspergillus nidulans; D005656:Fungal Proteins; D009566:Nitrates; D014157:Transcription Factors