Li Y, Flanagan P M, Tschochner H, Kornberg R D
Department of Cell Biology, Stanford University, School of Medicine, CA 94305.
Science. 1994 Feb 11;263(5148):805-7. doi: 10.1126/science.8303296.
An RNA polymerase II transcription system was resolved and reconstituted from extracts of Schizosaccharomyces pombe. Exchange with components of a Saccharomyces cerevisiae system was undertaken to reveal the factor or factors responsible for the difference in location of the transcription start site, about 30 base pairs and 40 to 120 base pairs downstream of the TATA box in S. pombe and S. cerevisiae, respectively. Two components, counterparts of human transcription factor IIF (TFIIF) and TFIIH, could be exchanged individually between systems without effect on the start site. Three components, counterparts of human TFIIB, TFIIE, and RNA polymerase II, could not be exchanged individually but could be swapped in the pairs TFIIE-TFIIH and TFIIB-RNA polymerase II, which demonstrates that there are functional interactions between these components. Moreover, exchange of the latter pair shifted the starting position, which shows that TFIIB and RNA polymerase II are solely responsible for determining the start site of transcription.
从粟酒裂殖酵母提取物中解析并重构了一种RNA聚合酶II转录系统。与酿酒酵母系统的组分进行交换，以揭示导致转录起始位点位置差异的一个或多个因子。在粟酒裂殖酵母和酿酒酵母中，转录起始位点分别位于TATA框下游约30个碱基对以及40至120个碱基对处。人转录因子IIF（TFIIF）和TFIIH的对应物这两个组分可以在系统之间单独交换，而不会影响起始位点。人TFIIB、TFIIE和RNA聚合酶II的对应物这三个组分不能单独交换，但可以在TFIIE-TFIIH和TFIIB-RNA聚合酶II这两对中进行交换，这表明这些组分之间存在功能相互作用。此外，后一对的交换改变了起始位置，这表明TFIIB和RNA聚合酶II单独负责确定转录起始位点。
