CDK12 全局性地刺激 RNA 聚合酶 II 转录延伸和羧基末端结构域磷酸化。

CDK12 globally stimulates RNA polymerase II transcription elongation and carboxyl-terminal domain phosphorylation.

机构信息

Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, UK.

Max Planck Institute for Biophysical Chemistry, Department of Molecular Biology, Am Fassberg 11, 37077 Göttingen, Germany.

出版信息

Nucleic Acids Res. 2020 Aug 20;48(14):7712-7727. doi: 10.1093/nar/gkaa514.

DOI:10.1093/nar/gkaa514

PMID:32805052

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7641311/

Abstract

Cyclin-dependent kinase 12 (CDK12) phosphorylates the carboxyl-terminal domain (CTD) of RNA polymerase II (pol II) but its roles in transcription beyond the expression of DNA damage response genes remain unclear. Here, we have used TT-seq and mNET-seq to monitor the direct effects of rapid CDK12 inhibition on transcription activity and CTD phosphorylation in human cells. CDK12 inhibition causes a genome-wide defect in transcription elongation and a global reduction of CTD Ser2 and Ser5 phosphorylation. The elongation defect is explained by the loss of the elongation factors LEO1 and CDC73, part of PAF1 complex, and SPT6 from the newly-elongating pol II. Our results indicate that CDK12 is a general activator of pol II transcription elongation and indicate that it targets both Ser2 and Ser5 residues of the pol II CTD.

摘要

周期蛋白依赖性激酶 12（CDK12）可使 RNA 聚合酶 II（pol II）羧基末端结构域（CTD）磷酸化，但它在 DNA 损伤反应基因表达之外的转录中的作用尚不清楚。在这里，我们使用 TT-seq 和 mNET-seq 来监测快速 CDK12 抑制对人类细胞中转录活性和 CTD 磷酸化的直接影响。CDK12 抑制会导致转录延伸的全基因组缺陷，以及 CTD Ser2 和 Ser5 磷酸化的整体减少。延伸缺陷可以通过从新延伸的 pol II 中丢失延伸因子 LEO1 和 CDC73（PAF1 复合物的一部分）和 SPT6 来解释。我们的结果表明，CDK12 是 pol II 转录延伸的通用激活剂，并表明它靶向 pol II CTD 的 Ser2 和 Ser5 残基。