Hahn S, Buratowski S, Sharp P A, Guarente L
Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
Cell. 1989 Sep 22;58(6):1173-81. doi: 10.1016/0092-8674(89)90515-1.
We report the cloning of the gene that encodes the yeast TATA binding protein TFIID. TFIID contains 240 amino acids and has no obvious sequence similarity to other known proteins. TFIID was synthesized in vitro and in two separate assays behaved identically to the protein purified from yeast. TFIID bound to TATA elements from the adenovirus major late promoter (TATAAAA) and the yeast LEU2 promoter (TATTTAA) and formed protein-DNA complexes stable to electrophoresis only in the presence of TFIIA. In vitro-synthesized yeast TFIID also complemented a mammalian in vitro transcription system that lacked TFIID. Comparison of the yeast TFIID gene with the yeast SPT15 gene (suppressor of Ty element insertions) showed that the two genes are identical. This finding indicates that the yeast TFIID activity defined in vitro is responsible for specific transcription in vivo.
我们报道了编码酵母TATA结合蛋白TFIID的基因的克隆。TFIID含有240个氨基酸,与其他已知蛋白质没有明显的序列相似性。TFIID在体外合成,在两项独立的测定中,其行为与从酵母中纯化的蛋白质完全相同。TFIID与腺病毒主要晚期启动子(TATAAAA)和酵母LEU2启动子(TATTTAA)的TATA元件结合,并且仅在TFIIA存在的情况下形成对电泳稳定的蛋白质-DNA复合物。体外合成的酵母TFIID也补充了缺乏TFIID的哺乳动物体外转录系统。将酵母TFIID基因与酵母SPT15基因(Ty元件插入的抑制子)进行比较,结果表明这两个基因是相同的。这一发现表明,体外定义的酵母TFIID活性负责体内的特异性转录。