Arcangioli B, Lescure B
Eur J Biochem. 1986 Feb 17;155(1):69-75. doi: 10.1111/j.1432-1033.1986.tb09459.x.
Yeast RNA polymerase II initiates in vitro transcription at two sites located within the vector DNA and the cloned promoter, on a recombinant plasmid DNA containing the yeast iso1 cytochrome c promoter. Both initiation sites are found within a DNA fragment hypersensitive to osmium tetroxide modification. Using a series of yeast iso1 cytochrome c promoter deletions, we have characterized an upstream DNA sequence required for optimal transcription from this site and shown in this case a correlation between osmium sensitivity and the capacity of RNA polymerase to initiate. However, perturbation of the double helix is not sufficient to generate a transcription initiation site. Insertion of 28 alternating AT residues at the EcoRV site of pBR322 generates an site hypersensitive to osmium tetroxide modification, that does not serve as a transcription start site.
酵母RNA聚合酶II在含有酵母异1细胞色素c启动子的重组质粒DNA上，于载体DNA和克隆启动子内的两个位点起始体外转录。两个起始位点都位于对四氧化锇修饰敏感的DNA片段内。通过一系列酵母异1细胞色素c启动子缺失实验，我们鉴定了该位点最佳转录所需的上游DNA序列，并表明在这种情况下，四氧化锇敏感性与RNA聚合酶起始能力之间存在相关性。然而，双螺旋的扰动不足以产生转录起始位点。在pBR322的EcoRV位点插入28个交替的AT残基会产生一个对四氧化锇修饰敏感的位点，但它不是转录起始位点。
