Department of Biochemistry and Molecular Genetics, University of Virginia Health System, Charlottesville, Virginia 22908, USA.
Genome Res. 2010 Dec;20(12):1679-88. doi: 10.1101/gr.109504.110. Epub 2010 Sep 20.
TATA-binding protein (TBP) nucleates the assembly of the transcription preinitiation complex (PIC), and although TBP can bind promoters with high stability in vitro, recent results establish that virtually the entire TBP population is highly dynamic in yeast nuclei in vivo. This dynamic behavior is surprising in light of models that posit that a stable TBP-containing scaffold facilitates transcription reinitiation at active promoters. The dynamic behavior of TBP is a consequence of the enzymatic activity of the essential Snf2/Swi2 ATPase Mot1, suggesting that ensuring a highly mobile TBP population is critical for transcriptional regulation on a global scale. Here high-resolution tiling arrays were used to define how perturbed TBP dynamics impact the precision of RNA synthesis in Saccharomyces cerevisiae. We find that Mot1 plays a broad role in establishing the precision and efficiency of RNA synthesis: In mot1-42 cells, RNA length changes were observed for 713 genes, about twice the number observed in set2Δ cells, which display a previously reported propensity for spurious initiation within open reading frames. Loss of Mot1 led to both aberrant transcription initiation and termination, with prematurely terminated transcripts representing the largest class of events. Genetic and genomic analyses support the conclusion that these effects on RNA length are mechanistically tied to dynamic TBP occupancies at certain types of promoters. These results suggest a new model whereby dynamic disassembly of the PIC can influence productive RNA synthesis.
TATA 结合蛋白(TBP)为转录起始前复合物(PIC)的组装提供核心作用,尽管 TBP 在体外可以高稳定性地结合启动子,但最近的研究结果表明,在酵母核内,实际上整个 TBP 群体都具有高度的动态性。鉴于某些模型认为稳定的 TBP 支架有助于在活性启动子处进行转录重新起始,这种动态行为令人惊讶。TBP 的动态行为是必需的 Snf2/Swi2 ATPase Mot1 的酶活性的结果,这表明确保高度移动的 TBP 群体对于全局转录调控至关重要。在这里,使用高分辨率平铺阵列来定义 TBP 动力学的扰动如何影响酿酒酵母中 RNA 合成的精度。我们发现 Mot1 在建立 RNA 合成的精度和效率方面发挥着广泛的作用:在 mot1-42 细胞中,观察到 713 个基因的 RNA 长度发生变化,大约是在 set2Δ 细胞中观察到的两倍,后者显示出在开放阅读框内发生虚假起始的先前报道的倾向。Mot1 的缺失导致转录起始和终止异常,过早终止的转录本代表最大的事件类别。遗传和基因组分析支持这样的结论,即这些对 RNA 长度的影响在机制上与某些类型启动子处的 TBP 动态占有率有关。这些结果表明了一个新的模型,即 PIC 的动态拆卸可以影响有效的 RNA 合成。