Analysis of Cre1 binding sites in the Trichoderma reesei cbh1 upstream region
Analysis of Cre1 binding sites in the Trichoderma reesei cbh1 upstream region
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DOI：
10.1016/s0378-1097(96)00432-6
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发表时间：
1996-12-15
影响因子：
2.1
通讯作者：
Uozumi, T
中科院分区：
文献类型：
--
作者：
Takashima, S;Iikura, H;Uozumi, T
对含有里氏木霉纤维二糖水解酶I基因(CBHL)上游区域的1.5kb XbaI-SacII片段进行了序列测定。1.5kb的片段包含8个6bp的位点，它们的序列与与分解代谢抑制因子CREA结合的共同序列相同或相似。与麦芽糖结合蛋白：：Cre1(10-131)融合蛋白(Cre1是里氏木霉的分解代谢抑制物)和cbh1上游区域的结合分析结果表明，在凝胶迁移率分析中，504bp的XbaI-NSPV片段(核苷酸位置-1496至-993)和356 bp的NSPV-muni片段(核苷酸位置-994至-639)发生了位移，其中XbaI-NSPV片段含有A1、A2和A3三个6bp位点。DNase I足迹实验表明，6个碱基的A2、B1、B2和B3位置不受DNase I消化的影响。
A 1.5-kb XbaI-SacII fragment containing the upstream region of the Trichoderma reesei cellobiohydrolase I gene (cbhl) 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.