CDK13 cooperates with CDK12 to control global RNA polymerase II processivity
CDK13 cooperates with CDK12 to control global RNA polymerase II processivity
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DOI：
10.1126/sciadv.aaz5041
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发表时间：
2020-04-01
期刊：
影响因子：
13.6
通讯作者：
Johnstone, Ricky W.
中科院分区：
文献类型：
--
作者：
Fan, Zheng;Devlin, Jennifer R.;Johnstone, Ricky W.
RNA聚合酶II（POLII）驱动的转录周期在不同的检查点受到细胞周期蛋白依赖性激酶（CDK）及其同源细胞周期蛋白的严格调控。支持转录延伸、持续合成能力和所涉及的CDK-细胞周期蛋白对的分子事件仍然知之甚少。使用CRISPR-Cas9同源定向修复，我们产生了CDK 12和CDK 13的类似物敏感性激酶变体，以探测它们各自的和共享的生物学和分子作用。CDK 12或CDK 13的单一抑制诱导与细胞生长信号传导途径和/或DNA损伤相关的转录应答，对细胞活力的影响最小。相反，双重激酶抑制有效地诱导细胞死亡，这与广泛的基因组范围的转录变化相关，包括广泛使用替代的3'多聚腺苷酸化位点。在分子水平上，双重激酶抑制导致POLII CTD磷酸化的丧失，并大大降低POLII延伸率和持续合成能力。这些数据定义了CDK 12和CDK 13之间的大量冗余，并将两者鉴定为全局POLII持续合成能力和转录延伸的基本调节因子。
The RNA polymerase II (POLII)-driven transcription cycle is tightly regulated at distinct checkpoints by cyclin-dependent kinases (CDKs) and their cognate cyclins. The molecular events underpinning transcriptional elongation, processivity, and the CDK-cyclin pair(s) involved remain poorly understood. Using CRISPR-Cas9 homology-directed repair, we generated analog-sensitive kinase variants of CDK1 2 and CDK13 to probe their individual and shared biological and molecular roles. Single inhibition of CDK12 or CDK13 induced transcriptional responses associated with cellular growth signaling pathways and/or DNA damage, with minimal effects on cell viability. In contrast, dual kinase inhibition potently induced cell death, which was associated with extensive genome-wide transcriptional changes including widespread use of alternative 3' polyadenylation sites. At the molecular level, dual kinase inhibition resulted in the loss of POLII CTD phosphorylation and greatly reduced POLII elongation rates and processivity. These data define substantial redundancy between CDK1 2 and CDK13 and identify both as fundamental regulators of global POLII processivity and transcription elongation.