Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK.
Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK.
Mol Cell. 2019 Apr 4;74(1):158-172.e9. doi: 10.1016/j.molcel.2019.01.027. Epub 2019 Feb 25.
The pervasive nature of RNA polymerase II (Pol II) transcription requires efficient termination. A key player in this process is the cleavage and polyadenylation (CPA) factor PCF11, which directly binds to the Pol II C-terminal domain and dismantles elongating Pol II from DNA in vitro. We demonstrate that PCF11-mediated termination is essential for vertebrate development. A range of genomic analyses, including mNET-seq, 3' mRNA-seq, chromatin RNA-seq, and ChIP-seq, reveals that PCF11 enhances transcription termination and stimulates early polyadenylation genome-wide. PCF11 binds preferentially between closely spaced genes, where it prevents transcriptional interference and consequent gene downregulation. Notably, PCF11 is sub-stoichiometric to the CPA complex. Low levels of PCF11 are maintained by an auto-regulatory mechanism involving premature termination of its own transcript and are important for normal development. Both in human cell culture and during zebrafish development, PCF11 selectively attenuates the expression of other transcriptional regulators by premature CPA and termination.
RNA 聚合酶 II（Pol II）转录的普遍性需要有效的终止。在这个过程中，一个关键的参与者是切割和多聚腺苷酸化（CPA）因子 PCF11，它直接与 Pol II C 末端结构域结合，并在体外将延伸的 Pol II 从 DNA 上拆除。我们证明了 PCF11 介导的终止对脊椎动物的发育是必不可少的。一系列的基因组分析，包括 mNET-seq、3' mRNA-seq、染色质 RNA-seq 和 ChIP-seq，揭示了 PCF11 增强转录终止并在全基因组范围内刺激早期多聚腺苷酸化。PCF11 优先结合在紧密间隔的基因之间，在那里它可以防止转录干扰和随后的基因下调。值得注意的是，PCF11 的数量低于 CPA 复合物。其自身转录物的过早终止涉及一个自动调节机制，可维持 PCF11 的低水平，这对正常发育很重要。在人类细胞培养和斑马鱼发育过程中，PCF11 通过过早的 CPA 和终止选择性地减弱其他转录调节剂的表达。
