Analysis of Cre1 binding sites in the Trichoderma reesei cbh1 upstream region.
A 1.5-kb XbaI-SacII fragment containing the upstream region of the Trichoderma reesei cellobiohydrolase I gene (cbh1) has been sequenced. The 1.5-kb fragment contains eight 6-bp sites having an identical or similar sequence to the consensus sequence for binding a catabolite repressor, Aspergillus nidulans CreA. Results of binding assays with the maltose-binding protein::Cre1(10-131) fusion protein (Cre1 is a catabolite repressor of T. reesei) and the cbh1 upstream region revealed that a 504-bp XbaI-NspV fragment (nucleotide position -1496 to -993) bearing three 6-bp sites, A1, A2, and A3, and a 356-bp NspV-MunI fragment (nucleotide position -994 to -639) bearing three 6-bp sites, B1, B2, and B3, were shifted in the electrophoretic mobility shift assay. DNase I footprinting experiments showed that the 6-bp sites A2, B1, B2, and B3 were protected from DNase I digestion.
Takashima
S
S
Department of Biotechnology, Graduate School of Agricultural and Life Sciences, University of Tokyo, Japan.
Iikura
H
H
Nakamura
A
A
Masaki
H
H
Uozumi
T
T
D001483:Base Sequence; D001665:Binding Sites / Q000502:physiology; D003001:Cloning, Molecular; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D003850:Deoxyribonuclease I; D005656:Fungal Proteins; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial / Q000235:genetics; D008969:Molecular Sequence Data; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D014242:Trichoderma / Q000235:genetics* / Q000378:metabolism; D016335:Zinc Fingers / Q000502:physiology
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D012097:Repressor Proteins; D003850:Deoxyribonuclease I