Witney F R, Surzycki J A, Seidman S, Dodds J R, Gussin G N, Surzycki S J
Mol Gen Genet. 1980;179(3):627-46. doi: 10.1007/BF00271753.
Electron microscopic visualization of binary complexes between eukaryotic RNA polymerases and Adenovirus 2 (Ad 2) DNA was used to locate specific binding sites for the enzymes. RNA polymerase II from human placenta binds to 10--16 distinct sites depending on the ratio of enzyme to DNA and the divalent cation present in the binding mixture. Wheat germ RNA polymerase binds to 12--14 strong binding sites and 2--3 weaker sites, all but one of which correspond to binding sites for the placental enzyme. At least six of the strong binding sites for both enzymes correspond to promoters known to be active in vivo. As a test of the two-state model for transcription initiation, we examined binding of wheat germ RNA polymerase II to Ad 2 DNA at 0 degrees and 37 degrees. The extent of binding was the same at the two temperatures and the distributions of binary complexes were virtually identical. This observation, in conjunction with results presented previously, is strong support for the existence of I and RS complexes in eukaryotic systems.
利用电子显微镜观察真核生物RNA聚合酶与腺病毒2(Ad 2)DNA之间的二元复合物,以定位这些酶的特异性结合位点。人胎盘来源的RNA聚合酶II根据酶与DNA的比例以及结合混合物中存在的二价阳离子,可结合到10 - 16个不同的位点。小麦胚芽RNA聚合酶可结合到12 - 14个强结合位点和2 - 3个较弱的位点,其中除一个位点外,其余位点均与胎盘酶的结合位点相对应。两种酶的至少六个强结合位点对应于已知在体内具有活性的启动子。作为对转录起始的双态模型的测试,我们检测了小麦胚芽RNA聚合酶II在0℃和37℃下与Ad 2 DNA的结合情况。在这两个温度下结合程度相同,二元复合物的分布实际上也相同。这一观察结果与先前呈现的结果相结合,有力地支持了真核生物系统中存在I和RS复合物。