Kitajima S, Kawaguchi T, Yasukochi Y, Weissman S M
Department of Human Genetics, Tokyo Medical and Dental University, Japan.
Proc Natl Acad Sci U S A. 1989 Aug;86(16):6106-10. doi: 10.1073/pnas.86.16.6106.
Multiple protein factors from HeLa cells are necessary for the accurate initiation of transcription on minimal promoters in vitro. We have partially purified these factors by chromatographic methods. In addition to RNA polymerase II, six factors A-F (FA, FB, FC, FD, FE, and FF) necessary for initiation at the beta-globin promoter start site in vitro have been identified. Certain of these (FA, FC, and FE) have been purified to near homogeneity. The present purification scheme yields sufficient amounts of purified material for the more detailed characterization and cloning of the genes for these activities. Among these factors, FD and FF were required with template DNA at an early step of formation of the initiation complex, whereas FB, FA together with FC, and FE were effective when added at successively later stages in the process of complex formation.
来自海拉细胞的多种蛋白质因子对于体外在最小启动子上准确起始转录是必需的。我们已经通过色谱方法部分纯化了这些因子。除了RNA聚合酶II外，还鉴定出了体外在β-珠蛋白启动子起始位点起始转录所需的六个因子A - F（FA、FB、FC、FD、FE和FF）。其中某些因子（FA、FC和FE）已纯化至接近均一性。目前的纯化方案产生了足够量的纯化材料，用于对这些活性的基因进行更详细的表征和克隆。在这些因子中，FD和FF在起始复合物形成的早期阶段需要与模板DNA一起存在，而FB、FA与FC以及FE在复合物形成过程中相继较晚阶段添加时才有效。
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