Characterization of the Aspergillus parasiticus major nitrogen regulatory gene, areA

Biochim Biophys Acta. 2000 Apr 25;1491(1-3):263-6. doi: 10.1016/s0167-4781(00)00004-x.

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.

MeSH terms

  • Aspergillus / genetics*
  • Base Sequence
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Fungal Proteins / biosynthesis
  • Fungal Proteins / chemistry
  • Fungal Proteins / genetics*
  • Nitrates / metabolism
  • Transcription Factors / biosynthesis
  • Transcription Factors / chemistry
  • Transcription Factors / genetics*
  • Transcription, Genetic

Substances

  • AreA protein, Aspergillus nidulans
  • Fungal Proteins
  • Nitrates
  • Transcription Factors