A novel nuclear factor, SREB, binds to a cis-acting element, SRE, required for inducible expression of the Aspergillus oryzae Taka-amylase A gene in A. nidulans.
The Taka-amylase A gene (taaG2) of Aspergillus oryzae is inducibly expressed in A. nidulans upon exposure to inducing carbon sources, such as starch and maltose. In order to identify nuclear factor(s) possibly involved in the induction of the taaG2 gene, gel mobility shift assays and DNase I footprinting analyses were carried out, and revealed a novel nuclear factor in A. nidulans extracts, which specifically bound to two sites in the taaG2 promoter region, -204 to -189 and -182 to -168, which share the common sequence GGAAATT. The nuclear factor was detected in nuclei from both induced and uninduced mycelia. Mutational analysis within and around the binding sequences demonstrated that only the upstream binding sequence, designated SRE (starch responsive element), was required for the inducible expression of the taaG2 gene, and thus we designated the nuclear factor SREB (SRE binding factor). The downstream binding site contained an inverted SRE (ISRE) and played no role in the induction of taaG2 expression. SREB was shown by gel retardation assays to have higher affinity for SRE than for ISRE.
Tani
S
S
Department of Biological Mechanisms and Functions, Graduate School of Bioagricultural Sciences, Nagoya University, Aichi, Japan.
Kawaguchi
T
T
Kato
M
M
Kobayashi
T
T
Tsukagoshi
N
N
D001233:Aspergillus nidulans / Q000378:metabolism; D001236:Aspergillus oryzae / Q000201:enzymology; D001483:Base Sequence; D018983:DNA Footprinting; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000502:physiology*; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D004926:Escherichia coli; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009687:Nuclear Proteins / Q000235:genetics* / Q000502:physiology*; D012689:Sequence Homology, Nucleic Acid; D000516:alpha-Amylases / Q000235:genetics* / Q000378:metabolism
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D000516:alpha-Amylases