TFIIH kinase CDK7 drives cell proliferation through a common core transcription factor network.. TFIIH kinase CDK7 drives cell proliferation through a common core transcription factor network.

We combined cryo-EM, transcriptomics, and a clinically relevant inhibitor to define how the TFIIH-associated CDK7 kinase coordinately controls the cell cycle and RNA polymerase II (RNAPII) transcription. CDK7 inhibition rapidly blocked expression of constitutively active genes, whereas inducible genes were unaffected. Distinct sets of sequence-specific, DNA-binding transcription factors (TFs) regulate constitutive vs. inducible genes; accordingly, inducible TFs (e.g. HSF1) were refractory to CDK7 inhibition. By contrast, CDK7 was required to maintain activity of a core set of promoter-associated TFs that drive proliferative gene expression programs; these core TFs (n=78) are constitutively active in proliferating cells. Thus, a major biological function for CDK7 is regulation of TFs that drive cell proliferation, revealing an apparent universal mechanism by which CDK7 coordinates RNAPII transcription with cell cycle regulation. Overall design: Precision run-on sequencing (PRO-seq) in HCT116 cells to evaluate the effects of CDK7 inhibition under basal and IFN-gamma stimulated conditions. There are 2 biological replicates for each condition.

我们结合冷冻电镜（cryo-EM）、转录组学与临床相关抑制剂，阐明了结合通用转录因子IIH（TFIIH）的CDK7激酶如何协同调控细胞周期与RNA聚合酶II（RNAPII）转录。CDK7抑制可快速阻断组成型活性基因的表达，而诱导型基因则不受影响。组成型基因与诱导型基因分别由不同的序列特异性DNA结合转录因子（TFs）调控；因此，诱导型转录因子（如热休克因子1（HSF1））对CDK7抑制不敏感。与之相反，CDK7对于维持一组驱动增殖基因表达程序的核心启动子结合转录因子的活性是必需的；这78个核心转录因子在增殖细胞中呈组成型激活状态。因此，CDK7的主要生物学功能之一是调控驱动细胞增殖的转录因子，进而揭示了CDK7协调RNAPII转录与细胞周期调控的潜在通用机制。整体实验设计：在HCT116细胞中开展精准延伸测序（PRO-seq），以评估CDK7抑制在基础培养条件与干扰素γ（IFN-γ）刺激条件下的调控效应。每个实验条件均设置2个生物学重复。