Heberlein U, England B, Tjian R
Cell. 1985 Jul;41(3):965-77. doi: 10.1016/s0092-8674(85)80077-5.
Fractionation of a nuclear extract derived from Drosophila tissue culture cells reveals the presence of multiple components involved in accurate transcription of both distal and proximal promoters of the alcohol dehydrogenase (Adh) gene. Transcription of deletion mutants indicates that a region between -24 and -85 upstream of the distal start site contains sequences required for RNA synthesis in vitro. Moreover, sequences that overlap this same upstream control region are specifically bound and protected from DNAase digestion by a promoter-specific transcription factor, Adf-1. Analysis of proximal promoter mutants identified multiple upstream elements that influence transcription, and DNAase footprint analysis detected three specific binding regions. Adf-1 binds at least one of these proximal promoter regions but interaction at this site is not specifically required for transcription. Our results suggest that multiple sequence-specific DNA binding proteins interact differentially with the proximal and distal promoters of Adh to activate transcription.
对源自果蝇组织培养细胞的核提取物进行分级分离，结果显示存在多种参与乙醇脱氢酶（Adh）基因远端和近端启动子精确转录的成分。缺失突变体的转录表明，远端起始位点上游-24至-85之间的区域包含体外RNA合成所需的序列。此外，与该相同上游控制区域重叠的序列被启动子特异性转录因子Adf-1特异性结合并保护免受DNA酶消化。近端启动子突变体的分析确定了多个影响转录的上游元件，DNA酶足迹分析检测到三个特异性结合区域。Adf-1结合这些近端启动子区域中的至少一个，但该位点的相互作用并非转录所必需。我们的结果表明，多种序列特异性DNA结合蛋白与Adh的近端和远端启动子发生不同的相互作用以激活转录。
