Flanagan P M, Kelleher R J, Tschochner H, Sayre M H, Kornberg R D
Department of Cell Biology, Fairchild Center, Stanford University School of Medicine, CA 94305.
Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7659-63. doi: 10.1073/pnas.89.16.7659.
Resolution of whole cell extract through two chromatographic steps yields a single protein fraction requiring only the addition of TFIID for the initiation of transcription at RNA polymerase II promoters. This approach allows the convenient generation of RNA polymerase II transcription systems from Saccharomyces cerevisiae, human lymphocytes, and Schizosaccharomyces pombe. TFIIDs from all three organisms are interchangeable among all three systems. The S. cerevisiae and Sch. pombe systems support effects of acidic activator proteins, provided a further protein fraction from S. cerevisiae is supplied. This further fraction is distinct from the mediator of transcriptional activation described previously and represents a second component in addition to general initiation factors that may facilitate a response to acidic activators.
通过两步色谱法对全细胞提取物进行分离，可得到单一蛋白质组分，该组分仅需添加TFIID即可在RNA聚合酶II启动子处起始转录。这种方法能够方便地从酿酒酵母、人淋巴细胞和粟酒裂殖酵母中生成RNA聚合酶II转录系统。来自这三种生物体的TFIID在所有这三个系统中均可互换。酿酒酵母和粟酒裂殖酵母系统支持酸性激活蛋白的作用，前提是提供来自酿酒酵母的另一蛋白质组分。这一另外的组分不同于先前描述的转录激活介质，并且代表了除一般起始因子之外的第二种组分，其可能有助于对酸性激活剂作出反应。
