Sci Adv . 2020 Apr 29
Vittorio Branchi, Tobias J Weismüller, Taotao Zhou, Jonas Henn, Alexander Semaan, Tim R Glowka, Maria Gonzalez-Carmona, Christian Strassburg, Jörg C Kalff, Steffen Manekeller, Hanno Matthaei, Hiroki Kato, Hideo Nagatomo, Mitsuo Nakai, Tatsuhiro Sakaiya, Hidenori Terasaki, Tadashi Kondo, Yoichiro Hironaka, Katsuya Shimizu, Keisuke Shigemori, Nasim Shahidi Hamedani, Jens Müller, Fabian Tolle, Heiko Rühl, Behnaz Pezeshkpoor, Kerstin Liphardt, Johannes Oldenburg, Günter Mayer, Bernd Pötzsch, Evelyn Gaffal, Andrea M Kemter, Stefanie Scheu, Rafael Leite Dantas, Jens Vogt, Bernhard Baune, Thomas Tüting, Andreas Zimmer, Judith Alferink, Florian Langer, Stefan Kluge, Robert Klamroth, Johannes Oldenburg, Anne L Fletcher, Alfie T Baker, Veronika Lukacs-Kornek, Konstantin Knoblich, K John Pasi, Kathelijn Fischer, Margaret Ragni, Roshni Kulkarni, Margareth C Ozelo, Johnny Mahlangu, Amy Shapiro, Stephanie P'Ng, Hervé Chambost, Beatrice Nolan, Carolyn Bennett, Tadashi Matsushita, Bent Winding, Joachim Fruebis, Huixing Yuan, Dan Rudin, Johannes Oldenburg, Huijun Wang, Aytaj Humbatova, Yuanxiang Liu, Wen Qin, Mingyang Lee, Nicole Cesarato, Fanny Kortüm, Sheetal Kumar, Maria Teresa Romano, Shangzhi Dai, Ran Mo, Sugirthan Sivalingam, Susanne Motameny, Yuan Wu, Xiaopeng Wang, Xinwu Niu, Songmei Geng, Dorothea Bornholdt, Peter M Kroisel, Gianluca Tadini, Scott D Walter, Fabian Hauck, Katta M Girisha, Anne-Marie Calza, Armand Bottani, Janine Altmüller, Andreas Buness, Shuxia Yang, Xiujuan Sun, Lin Ma, Kerstin Kutsche, Karl-Heinz Grzeschik, Regina C Betz, Zhimiao Lin, Zheng Fan, Jennifer R Devlin, Simon J Hogg, Maria A Doyle, Paul F Harrison, Izabela Todorovski, Leonie A Cluse, Deborah A Knight, Jarrod J Sandow, Gareth Gregory, Andrew Fox, Traude H Beilharz, Nicholas Kwiatkowski, Nichollas E Scott, Ana Tufegdzic Vidakovic, Gavin P Kelly, Jesper Q Svejstrup, Matthias Geyer, Nathanael S Gray, Stephin J Vervoort, Ricky W Johnstone
The RNA polymerase II (POLII)-driven transcription cycle is tightly regulated at distinct checkpoints by cyclin-dependent kinases (CDKs) and their cognate cyclins. The molecular events underpinning transcriptional elongation, processivity, and the CDK-cyclin pair(s) involved remain poorly understood. Using CRISPR-Cas9 homology-directed repair, we generated analog-sensitive kinase variants of CDK12 and CDK13 to probe their individual and shared biological and molecular roles. Single inhibition of CDK12 or CDK13 induced transcriptional responses associated with cellular growth signaling pathways and/or DNA damage, with minimal effects on cell viability. In contrast, dual kinase inhibition potently induced cell death, which was associated with extensive genome-wide transcriptional changes including widespread use of alternative 3' polyadenylation sites. At the molecular level, dual kinase inhibition resulted in the loss of POLII CTD phosphorylation and greatly reduced POLII elongation rates and processivity. These data define substantial redundancy between CDK12 and CDK13 and identify both as fundamental regulators of global POLII processivity and transcription elongation.