Department of Biochemistry and Molecular Genetics, University of Virginia Health System, Charlottesville, VA, 22908, USA.
Center for Public Health Genomics, University of Virginia Health System, Charlottesville, VA, 22908, USA.
EMBO J. 2024 May;43(9):1799-1821. doi: 10.1038/s44318-024-00089-2. Epub 2024 Apr 2.
A great deal of work has revealed, in structural detail, the components of the preinitiation complex (PIC) machinery required for initiation of mRNA gene transcription by RNA polymerase II (Pol II). However, less-well understood are the in vivo PIC assembly pathways and their kinetics, an understanding of which is vital for determining how rates of in vivo RNA synthesis are established. We used competition ChIP in budding yeast to obtain genome-scale estimates of the residence times for five general transcription factors (GTFs): TBP, TFIIA, TFIIB, TFIIE and TFIIF. While many GTF-chromatin interactions were short-lived ( < 1 min), there were numerous interactions with residence times in the range of several minutes. Sets of genes with a shared function also shared similar patterns of GTF kinetic behavior. TFIIE, a GTF that enters the PIC late in the assembly process, had residence times correlated with RNA synthesis rates. The datasets and results reported here provide kinetic information for most of the Pol II-driven genes in this organism, offering a rich resource for exploring the mechanistic relationships between PIC assembly, gene regulation, and transcription.
大量的工作已经揭示了启动前复合物(PIC)机械所需的结构细节,这些复合物是 RNA 聚合酶 II(Pol II)启动 mRNA 基因转录所必需的。然而,对于体内 PIC 组装途径及其动力学的理解还不太清楚,对于确定体内 RNA 合成速率是如何建立的,这种理解是至关重要的。我们使用芽殖酵母中的竞争 ChIP 实验,获得了五个通用转录因子(GTFs):TBP、TFIIA、TFIIB、TFIIE 和 TFIIF 的基因组规模的停留时间估计值。虽然许多 GTF-染色质相互作用是短暂的(<1 分钟),但也有许多相互作用的停留时间在几分钟范围内。具有共同功能的基因集也具有相似的 GTF 动力学行为模式。TFIIE 是一种在组装过程后期进入 PIC 的 GTF,其停留时间与 RNA 合成速率相关。这里报告的数据集和结果为该生物中大多数 Pol II 驱动的基因提供了动力学信息,为探索 PIC 组装、基因调控和转录之间的机制关系提供了丰富的资源。