TY - JOUR

T1 - CDK13 cooperates with CDK12 to control global RNA polymerase II processivity

AU - Fan, Zheng

AU - Devlin, Jennifer R.

AU - Hogg, Simon J.

AU - Doyle, Maria A.

AU - Harrison, Paul F.

AU - Todorovski, Izabela

AU - Cluse, Leonie A.

AU - Knight, Deborah A.

AU - Sandow, Jarrod J.

AU - Gregory, Gareth

AU - Fox, Andrew

AU - Beilharz, Traude H.

AU - Kwiatkowski, Nicholas

AU - Scott, Nichollas E.

AU - Vidakovic, Ana Tufegdzic

AU - Kelly, Gavin P.

AU - Svejstrup, Jesper Q.

AU - Geyer, Matthias

AU - Gray, Nathanael S.

AU - Vervoort, Stephin J.

AU - Johnstone, Ricky W.

N1 - Funding Information: This work was supported by a project grant (to R.W.J.) and fellowship (to S.J.H.) from the Cancer Council Victoria, project grant support from the NHMRC (to R.W.J.), NHMRC Program (grant 454569 to R.W.J.), NHMRC Senior Principal Research Fellowship (to R.W.J.), Victorian Cancer Agency Early Career Seed Grant (to J.R.D.), Melbourne Research Scholarship (to Z.F.), and The Kids' Cancer Project (to R.W.J. and S.J.V.). S.J.V. was supported by a Rubicon Fellowship from the Netherlands Organization for Scientific Research (NWO, 019.161LW.017). We acknowledge support from the Peter MacCallum Cancer Centre Foundation, the Australian Cancer Research Foundation, and the Victorian Government's Operational Infrastructure Support Program. Work in the Svejstrup laboratory was supported by the Francis Crick Institute, which receives its core funding from Cancer Research UK (FC001166), the UK Medical Research Council (FC001166), and the Wellcome Trust (FC001166). Publisher Copyright: Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

PY - 2020

Y1 - 2020

N2 - 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.

AB - 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.

U2 - 10.1126/sciadv.aaz5041

DO - 10.1126/sciadv.aaz5041

M3 - Journal article

C2 - 32917631

AN - SCOPUS:85084667579

VL - 6

JO - Science advances

JF - Science advances

SN - 2375-2548

IS - 18

M1 - eaaz5041

ER -