Aspergillus nidulans nuclear proteins bind to a CCAAT element and the adjacent upstream sequence in the promoter region of the starch-inducible Taka-amylase A gene.
Aspergillus nidulans was used as an intermediate host to investigate the regulation of the Taka-amylase A (TAA) gene from Aspergillus oryzae. The induction of Taa by starch was confirmed to be regulated at the transcriptional level by analyzing the transcripts specific for Taa synthesized in vitro in nuclei from starch- and glucose-grown cells. A 55 bp DNA fragment containing a consensus CCAAT sequence from the promoter region of the Taa gene was shown to confer starch inducibility on the gene. A nuclear extract from starch-grown cells was assayed for proteins which bind to the promoter region of the Taa gene. A protein designated AnCP1 bound to the CCAAT sequence. A nuclear extract from glucose-grown cells contained two DNA-binding proteins designated AnCP2 and AnNP1. AnCP2 bound to the same CCAAT sequence as AnCP1, while AnNP1 bound to the 25 bp region just upstream of the AnCP2 binding site. Occupancy of the two binding sites appeared to be mutually exclusive, which is suggestive of a negative regulatory mechanism for gene expression.
Nagata
O
O
Department of Food Science and Technology, Faculty of Agriculture, Nagoya University, Japan.
Takashima
T
T
Tanaka
M
M
Tsukagoshi
N
N
D001233:Aspergillus nidulans / Q000235:genetics*; D001483:Base Sequence; D022762:CCAAT-Enhancer-Binding Proteins; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000378:metabolism; D016384:Consensus Sequence; D004268:DNA-Binding Proteins / Q000378:metabolism; D004790:Enzyme Induction / Q000187:drug effects; D005656:Fungal Proteins / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal / Q000235:genetics*; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012331:RNA, Fungal / Q000096:biosynthesis; D013213:Starch / Q000494:pharmacology; D013329:Structure-Activity Relationship; D000516:alpha-Amylases / Q000235:genetics*
D022762:CCAAT-Enhancer-Binding Proteins; C080284:CP1 protein, Aspergillus nidulans; C080285:CP2 protein, Aspergillus nidulans; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C080286:NP1 protein, Aspergillus nidulans; D009687:Nuclear Proteins; D012331:RNA, Fungal; D013213:Starch; D000516:alpha-Amylases