Sequence-specific binding sites in the Taka-amylase A G2 promoter for the CreA repressor mediating carbon catabolite repression.
The N-terminal part of the CreA protein encompassing two zinc fingers was expressed in Escherichia coli as a fusion protein with the maltose binding protein (MalE) of E. coli. Our results show that CreA binds to the promoter of the Taa-G2 gene encoding Taka-amylase A of Aspergillus oryzae. DNase I footprinting experiments showed that CreA bound to three sites with high affinity and to one site with low affinity within the first 401-bp region upstream of the transcription initiation site. All of the sites contained sequences related to the CreA consensus binding site (5'-SYGGRG-3'), and are suggested to participate in repression of the Taa-G2 gene in response to glucose.
Kato
M
M
Department of Applied Biological Sciences, Faculty of Agriculture, Nagoya University, Japan.
Sekine
K
K
Tsukagoshi
N
N
D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D002244:Carbon / Q000378:metabolism*; D002850:Chromatography, Gel; D018983:DNA Footprinting; D003850:Deoxyribonuclease I / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D015183:Restriction Mapping; D016335:Zinc Fingers / Q000235:genetics; D000516:alpha-Amylases / Q000235:genetics / Q000378:metabolism*
D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; C071237:CreA protein, Aspergillus nidulans; D002244:Carbon; D003850:Deoxyribonuclease I; D000516:alpha-Amylases