Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, United States.
Elife. 2021 Sep 13;10:e67964. doi: 10.7554/eLife.67964.
The preinitiation complex (PIC) for transcriptional initiation by RNA polymerase (Pol) II is composed of general transcription factors that are highly conserved. However, analysis of ChIP-seq datasets reveals kinetic and compositional differences in the transcriptional initiation process among eukaryotic species. In yeast, Mediator associates strongly with activator proteins bound to enhancers, but it transiently associates with promoters in a form that lacks the kinase module. In contrast, in human, mouse, and fly cells, Mediator with its kinase module stably associates with promoters, but not with activator-binding sites. This suggests that yeast and metazoans differ in the nature of the dynamic bridge of Mediator between activators and Pol II and the composition of a stable inactive PIC-like entity. As in yeast, occupancies of TATA-binding protein (TBP) and TBP-associated factors (Tafs) at mammalian promoters are not strictly correlated. This suggests that within PICs, TFIID is not a monolithic entity, and multiple forms of TBP affect initiation at different classes of genes. TFIID in flies, but not yeast and mammals, interacts strongly at regions downstream of the initiation site, consistent with the importance of downstream promoter elements in that species. Lastly, Taf7 and the mammalian-specific Med26 subunit of Mediator also interact near the Pol II pause region downstream of the PIC, but only in subsets of genes and often not together. Species-specific differences in PIC structure and function are likely to affect how activators and repressors affect transcriptional activity.
转录起始前复合物 (PIC) 由 RNA 聚合酶 (Pol) II 的通用转录因子组成,这些因子高度保守。然而,对 ChIP-seq 数据集的分析表明,真核生物物种之间的转录起始过程在动力学和组成上存在差异。在酵母中,中介体与结合增强子的激活蛋白强烈结合,但它以缺乏激酶模块的形式与启动子短暂结合。相比之下,在人类、小鼠和果蝇细胞中,具有激酶模块的中介体与启动子稳定结合,但与激活剂结合位点不结合。这表明酵母和后生动物在中介体作为激活剂和 Pol II 之间的动态桥梁的性质以及稳定的无活性 PIC 样实体的组成上存在差异。与酵母一样,哺乳动物启动子上 TATA 结合蛋白 (TBP) 和 TBP 相关因子 (Taf) 的占有率并不严格相关。这表明在 PIC 内,TFIID 不是一个整体实体,并且多种形式的 TBP 影响不同类别的基因的起始。在果蝇中,但不在酵母和哺乳动物中,TFIID 在起始位点下游区域强烈相互作用,这与该物种中下游启动子元件的重要性一致。最后,Taf7 和中介体的哺乳动物特异性 Med26 亚基也在 PIC 下游的 Pol II 暂停区域附近相互作用,但仅在部分基因中,并且通常不是一起作用。PIC 结构和功能的物种特异性差异可能会影响激活剂和抑制剂如何影响转录活性。