CTDP1 and RPB7 Stabilize Pol II and Permit Reinitiation

The mechanisms governing the termination and subsequent reinitiation of RNA polymerase II (Pol II) remain poorly understood. Here we found that depletion of RPB7 leads to the destabilization of Pol II’s largest subunit, RPB1. This destabilization is influenced by the loop regions of RPB7, CDK9, the C-terminal domain (CTD) of RPB1, and its linker region. The stabilization process of RPB1 is regulated by the E3 ubiquitin ligase Cullin 3. Additionally, RPB7 interacts with the phosphatase CTDP1, which is crucial for maintaining RPB1 stability. RPB7 is also vital for the reinitiation of Pol II, engages with RNA processing factors, and is localized to the RNA exit channel of the Pol II complex. The absence of RPB7 compromises RNA processing. We propose that RPB7 recruits CTDP1 to dephosphorylate Pol II, enhancing its stability and facilitating efficient reinitiation, adding a new dimension to transcriptional regulation. In this dataset, the raw data of Western blot, genotyping and imaging in our study was uploaded. The file name has the corresponding figure number in the paper with the detected protein's name (for Western blot) or its treatment condition. For cell imaging, images were acquired by Nikon A1R microscope. More details can be found in our paper.调控RNA聚合酶II（RNA polymerase II, Pol II）的终止及后续重新起始的分子机制目前仍不甚明晰。本研究发现，敲低RPB7会引发Pol II最大亚基RPB1的稳定性下降。该不稳定过程受RPB7、CDK9的环区，以及RPB1的C端结构域（C-terminal domain, CTD）及其连接区域调控。RPB1的稳定过程受E3泛素连接酶Cullin 3的调控。此外，RPB7可与磷酸酶CTDP1相互作用，该相互作用对维持RPB1的稳定性至关重要。RPB7对于Pol II的重新起始同样不可或缺，其可与RNA加工因子结合，并定位于Pol II复合物的RNA出口通道。缺失RPB7会损害RNA加工过程。我们提出，RPB7可招募CTDP1对Pol II进行去磷酸化修饰，进而增强其稳定性并促进高效的重新起始过程，为转录调控研究增添了新的维度。 本数据集上传了本研究中蛋白质免疫印迹（Western blot）、基因分型及成像实验的原始数据。文件名包含论文中对应的图号，对于蛋白质免疫印迹实验，文件名标注了检测蛋白的名称；对于其他实验，则标注了处理条件。细胞成像实验的图像均通过尼康A1R显微镜采集。更多细节可参见本研究论文。