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Cyclin-dependent kinase 7 (CDK7), part of the general transcription factor TFIIH, promotes gene transcription by phosphorylating the C-terminal domain of RNA polymerase II (RNA Pol II). Here, we combine rapid CDK7 kinase inhibition with multi-omics analysis to unravel the direct functions of CDK7 in human cells. CDK7 inhibition causes RNA Pol II retention at promoters, leading to decreased RNA Pol II initiation and immediate global downregulation of transcript synthesis. Elongation, termination, and recruitment of co-transcriptional factors are not directly affected. Although RNA Pol II, initiation factors, and Mediator accumulate at promoters, RNA Pol II complexes can also proceed into gene bodies without promoter-proximal pausing while retaining initiation factors and Mediator. Further downstream, RNA Pol II phosphorylation increases and initiation factors and Mediator are released, allowing recruitment of elongation factors and an increase in RNA Pol II elongation velocity. Collectively, CDK7 kinase activity promotes the release of initiation factors and Mediator from RNA Pol II, facilitating RNA Pol II escape from the promoter.

Original publication

DOI

10.1016/j.molcel.2024.05.007

Type

Journal article

Journal

Mol Cell

Publication Date

20/06/2024

Volume

84

Pages

2287 - 2303.e10

Keywords

CDK7, CTD, Mediator, RNA polymerase II, analog sensitive, gene regulation, pre-initiation complex, promoter escape, transcription, transcription initiation, Cyclin-Dependent Kinase-Activating Kinase, Humans, Cyclin-Dependent Kinases, RNA Polymerase II, Promoter Regions, Genetic, Phosphorylation, Transcription Initiation, Genetic, Protein Kinase Inhibitors, Mediator Complex, HeLa Cells, Transcription Factor TFIIH, HEK293 Cells