Gonzalez-Couto E, Klages N, Strubin M
Department of Genetics and Microbiology, University Medical Centre, Rue Michel Servet 1, 1211 Geneva 4, Switzerland.
Proc Natl Acad Sci U S A. 1997 Jul 22;94(15):8036-41. doi: 10.1073/pnas.94.15.8036.
Eukaryotic transcriptional activators may function by stimulating formation of RNA polymerase II preinitiation complexes at the core promoter of genes. In this case, their mode of action will intrinsically depend on how these complexes assemble on promoters in living cells, an issue that remains largely unexplored. Here we show that in yeast the basal transcription machinery is brought to the promoter in the form of at least two subcomplexes, TFIID and a complex comprising TFIIB and other essential components. Individual recruitment of either complex by artificial contact with a transcriptionally inactive, sequence-specific DNA-binding protein suffices to trigger transcriptional activation from a wild-type core promoter bearing the appropriate binding site. In contrast, activation from a promoter containing a weakened TATA element is only observed upon recruitment of TFIID. Tethering TFIIB on that promoter remains without effect, but the simultaneous recruitment of both components leads to strong synergistic activation. These findings suggest a simple mechanism whereby two activators that contact distinct subcomplexes of the basal machinery may stimulate transcription synergistically and differentially depending on the nature of the promoter.
真核转录激活因子可能通过刺激基因核心启动子处RNA聚合酶II预起始复合物的形成来发挥作用。在这种情况下,它们的作用方式本质上取决于这些复合物如何在活细胞的启动子上组装,而这一问题在很大程度上仍未得到探索。在这里,我们表明在酵母中,基础转录机制以至少两个亚复合物的形式被带到启动子上,即TFIID和一个包含TFIIB及其他必需成分的复合物。通过与转录无活性的、序列特异性DNA结合蛋白进行人工接触,单独招募这两种复合物中的任何一种,都足以从带有适当结合位点的野生型核心启动子触发转录激活。相比之下,只有在招募TFIID时,才能观察到从含有弱化TATA元件的启动子的激活。在该启动子上拴系TFIIB没有效果,但同时招募这两种成分会导致强烈的协同激活。这些发现提示了一种简单的机制,即两个与基础机制不同亚复合物接触的激活因子,可能根据启动子性质协同且有差异地刺激转录。