Department of Molecular Biology, Graduate School of Biomedical Sciences, Rowan University, Stratford, New Jersey, USA.
Department of Molecular Biology, Graduate School of Biomedical Sciences, Rowan University, Stratford, New Jersey, USA.
J Biol Chem. 2020 Nov 27;295(48):16280-16291. doi: 10.1074/jbc.RA120.015215. Epub 2020 Sep 15.
The Cdk8 kinase module (CKM) is a detachable Mediator subunit composed of cyclin C and one each of paralogs Cdk8/Cdk19, Med12/Med12L, and Med13/Med13L. Our previous RNA-Seq studies demonstrated that cyclin C represses a subset of hydrogen peroxide-induced genes under normal conditions but is involved in activating other loci following stress. Here, we show that cyclin C directs this transcriptional reprograming through changes in its promoter occupancy. Following peroxide stress, cyclin C promoter occupancy increased for genes it activates while decreasing at loci it represses under normal conditions. Promoter occupancy of other CKM components generally mirrored cyclin C, indicating that the CKM moves as a single unit. It has previously been shown that some cyclin C leaves the nucleus following cytotoxic stress to induce mitochondrial fragmentation and apoptosis. We observed that CKM integrity appeared compromised at a subset of repressed promoters, suggesting a source of cyclin C that is targeted for nuclear release. Interestingly, mTOR inhibition induced a new pattern of cyclin C promoter occupancy indicating that this control is fine-tuned to the individual stress. Using inhibitors, we found that Cdk8 kinase activity is not required for CKM movement or repression but was necessary for full gene activation. In conclusion, this study revealed that different stress stimuli elicit specific changes in CKM promoter occupancy correlating to altered transcriptional outputs. Finally, although CKM components were recruited or expelled from promoters as a unit, heterogeneity was observed at individual promoters, suggesting a mechanism to generate gene- and stress-specific responses.
CDK8 激酶模块(CKM)是一个可分离的 Mediator 亚基,由细胞周期蛋白 C 和一个 CDK8/Cdk19、Med12/Med12L 和 Med13/Med13L 的同源物组成。我们之前的 RNA-Seq 研究表明,细胞周期蛋白 C 在正常情况下抑制一组过氧化氢诱导的基因,但在应激后参与激活其他基因。在这里,我们表明细胞周期蛋白 C 通过改变其启动子占据来指导这种转录重编程。在过氧化物应激后,细胞周期蛋白 C 启动子占据增加了其激活的基因,而在正常条件下抑制的基因则减少。其他 CKM 成分的启动子占据通常与细胞周期蛋白 C 相似,表明 CKM 作为一个整体移动。先前已经表明,一些细胞周期蛋白 C 在细胞毒性应激后离开细胞核,以诱导线粒体片段化和细胞凋亡。我们观察到 CKM 的完整性在一部分被抑制的启动子上似乎受到了损害,这表明细胞周期蛋白 C 的一个靶点是针对核释放的。有趣的是,mTOR 抑制诱导了细胞周期蛋白 C 启动子占据的新模式,表明这种控制是针对个体应激进行微调的。使用抑制剂,我们发现 Cdk8 激酶活性不是 CKM 运动或抑制所必需的,但对于完全基因激活是必需的。总之,这项研究表明,不同的应激刺激会引起 CKM 启动子占据的特定变化,与改变的转录输出相关。最后,尽管 CKM 成分作为一个整体被招募或驱逐出启动子,但在单个启动子上观察到异质性,这表明了一种产生基因和应激特异性反应的机制。