Selective inhibition of CDK7 reveals high-confidence targets and new models for TFIIH function in transcription
| Autor: | Goran Malojčić, Martine Brault, Ines H. Kaltheuner, Lewis C. Cantley, Patrick Deroy, Heeyoun Bunch, Shanhu Hu, Janet Iwasa, Robin D. Dowell, Michael J. Bradley, Christian Clavette, Limei Tao, Peter W. White, Matthias Geyer, Zachary C. Poss, William M. Old, Zachary L. Maas, Shraddha Nayak, Tim-Michael Decker, Christopher C. Ebmeier, Benjamin Erickson, Leah J. Damon, David Bentley, Jenna K. Rimel, Tomer M. Yaron, Jason J. Marineau, Jared L. Johnson, Kristin B. Hamman, Dylan J. Taatjes |
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| Rok vydání: | 2020 |
| Předmět: |
Transcription
Genetic Cell Survival RNA polymerase II HL-60 Cells Biology Models Biological 03 medical and health sciences 0302 clinical medicine Transcription (biology) Genetics Humans 030304 developmental biology 0303 health sciences Kinase Cyclin-Dependent Kinases Cell biology Enzyme Activation Alternative Splicing 030220 oncology & carcinogenesis RNA splicing Transcription factor II H biology.protein Cyclin-dependent kinase 9 Cyclin-dependent kinase 7 Transcription Factor TFIIH Cyclin-Dependent Kinase-Activating Kinase Developmental Biology CDK12 Research Paper |
| Zdroj: | Genes Dev |
| ISSN: | 1549-5477 |
| Popis: | CDK7 associates with the 10-subunit TFIIH complex and regulates transcription by phosphorylating the C-terminal domain (CTD) of RNA polymerase II (RNAPII). Few additional CDK7 substrates are known. Here, using the covalent inhibitor SY-351 and quantitative phosphoproteomics, we identified CDK7 kinase substrates in human cells. Among hundreds of high-confidence targets, the vast majority are unique to CDK7 (i.e., distinct from other transcription-associated kinases), with a subset that suggest novel cellular functions. Transcription-associated factors were predominant CDK7 substrates, including SF3B1, U2AF2, and other splicing components. Accordingly, widespread and diverse splicing defects, such as alternative exon inclusion and intron retention, were characterized in CDK7-inhibited cells. Combined with biochemical assays, we establish that CDK7 directly activates other transcription-associated kinases CDK9, CDK12, and CDK13, invoking a “master regulator” role in transcription. We further demonstrate that TFIIH restricts CDK7 kinase function to the RNAPII CTD, whereas other substrates (e.g., SPT5 and SF3B1) are phosphorylated by the three-subunit CDK-activating kinase (CAK; CCNH, MAT1, and CDK7). These results suggest new models for CDK7 function in transcription and implicate CAK dissociation from TFIIH as essential for kinase activation. This straightforward regulatory strategy ensures CDK7 activation is spatially and temporally linked to transcription, and may apply toward other transcription-associated kinases. |
| Databáze: | OpenAIRE |
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