Abstract
Gene expression by RNA polymerase II (RNAPII) is tightly controlled by cyclin-dependent kinases (CDKs) at discrete checkpoints during the transcription cycle. The pausing checkpoint following transcription initiation is primarily controlled by CDK9. We discovered that CDK9-mediated, RNAPII-driven transcription is functionally opposed by a protein phosphatase 2A (PP2A) complex that is recruited to transcription sites by the Integrator complex subunit INTS6. PP2A dynamically antagonizes phosphorylation of key CDK9 substrates including DSIF and RNAPII-CTD. Loss of INTS6 results in resistance to tumor cell death mediated by CDK9 inhibition, decreased turnover of CDK9 phospho-substrates, and amplification of acute oncogenic transcriptional responses. Pharmacological PP2A activation synergizes with CDK9 inhibition to kill both leukemic and solid tumor cells, providing therapeutic benefit in vivo. These data demonstrate that fine control of gene expression relies on the balance between kinase and phosphatase activity throughout the transcription cycle, a process dysregulated in cancer that can be exploited therapeutically.
| Original language | English |
|---|---|
| Pages (from-to) | 3143-3162.E32 |
| Number of pages | 52 |
| Journal | Cell |
| Volume | 184 |
| Issue number | 12 |
| DOIs | |
| Publication status | Published - 10 Jun 2021 |
Keywords
- cancer
- CDK9
- CRISPR-Cas9 screen
- CTD
- Integrator
- pause-release
- phosphatase
- PP2A
- PP2A activation
- RNA polymerase II
- transcriptional elongation
Access to Document
Other files and links
Cite this
- APA
- Author
- BIBTEX
- Harvard
- Standard
- RIS
- Vancouver
}
In: Cell, Vol. 184, No. 12, 10.06.2021, p. 3143-3162.E32.
Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - The PP2A-Integrator-CDK9 axis fine-tunes transcription and can be targeted therapeutically in cancer
AU - Vervoort, Stephin J.
AU - Welsh, Sarah A.
AU - Devlin, Jennifer R.
AU - Barbieri, Elisa
AU - Knight, Deborah A.
AU - Offley, Sarah
AU - Bjelosevic, Stefan
AU - Costacurta, Matteo
AU - Todorovski, Izabela
AU - Kearney, Conor J.
AU - Sandow, Jarrod J.
AU - Fan, Zheng
AU - Blyth, Benjamin
AU - McLeod, Victoria
AU - Vissers, Joseph H.A.
AU - Pavic, Karolina
AU - Martin, Ben P.
AU - Gregory, Gareth
AU - Demosthenous, Elena
AU - Zethoven, Magnus
AU - Kong, Isabella Y.
AU - Hawkins, Edwin D.
AU - Hogg, Simon J.
AU - Kelly, Madison J.
AU - Newbold, Andrea
AU - Simpson, Kaylene J.
AU - Kauko, Otto
AU - Harvey, Kieran F.
AU - Ohlmeyer, Michael
AU - Westermarck, Jukka
AU - Gray, Nathanael
AU - Gardini, Alessandro
AU - Johnstone, Ricky W.
N1 - Funding Information: We thank members of the Johnstone and Gardini laboratories, the Translational Research Laboratory, the Victorian Centre for Functional Genomics (VCFG), Molecular Genomics Core and the Flow Cytometry Core Facilities at the Peter MacCallum Cancer Center (PMCC), the Proteomics Facility at the Walter and Eliza Hall Institute, and the Genomics Core and the Proteomics Core at The Wistar Institute (P30-CA010815) for their technical expertise and for providing reagents. We thank Prof. Robert Fisher for providing phospho-Spt5 antibodies. Illustrations were made with BioRender.com . R.W.J. was supported by the Cancer Council Victoria , National Health and Medical Research Council of Australia (NHMRC) and The Kids’ Cancer Project . The Gardini laboratory (A.G.) was supported the American Cancer Society ( RSG-18-157-01-DMC ), the G. Harold & Leila Y. Mathers Foundation (A.G.), the NIH ( R01 HL141326 ), the Emerson Collective Cancer Research Fund , and the Ovarian Cancer Research Alliance . S.J.V. was supported by a Rubicon fellowship (NWO, 019.161LW.017 ), NHMRC EL1 fellowship ( GNT1178339 ), and The Kids’ Cancer Project . S.A.W. was supported by a training grant from NIH ( T32-GM071339 ). J.R.D. was supported by an Early Career Seed Grant from the Victorian Cancer Agency (VCA). S.J.H. was supported by a Postdoctoral Fellowship from the Cancer Council of Victoria (CCV). K.F.H. was supported by a NHMRC Senior Research Fellowship ( APP1078220 ). J.W. and K.P. were supported by Academy of Finland ( 294850 ) and Finnish Cancer Foundation . K.P. was supported by Finnish Cultural Foundation . The VCFG ACRF Translational Reverse Phase Protein Array platform (K.J.S.) is funded by the Australian Cancer Research Foundation (ACRF), University of Melbourne Collaborative Research Infrastructure Program , and the PMCC Foundation . The PMCC Foundation and ACRF provide generous support for equipment and core facilities. Funding Information: We thank members of the Johnstone and Gardini laboratories, the Translational Research Laboratory, the Victorian Centre for Functional Genomics (VCFG), Molecular Genomics Core and the Flow Cytometry Core Facilities at the Peter MacCallum Cancer Center (PMCC), the Proteomics Facility at the Walter and Eliza Hall Institute, and the Genomics Core and the Proteomics Core at The Wistar Institute (P30-CA010815) for their technical expertise and for providing reagents. We thank Prof. Robert Fisher for providing phospho-Spt5 antibodies. Illustrations were made with BioRender.com. R.W.J. was supported by the Cancer Council Victoria, National Health and Medical Research Council of Australia (NHMRC) and The Kids? Cancer Project. The Gardini laboratory (A.G.) was supported the American Cancer Society (RSG-18-157-01-DMC), the G. Harold & Leila Y. Mathers Foundation (A.G.), the NIH (R01 HL141326), the Emerson Collective Cancer Research Fund, and the Ovarian Cancer Research Alliance. S.J.V. was supported by a Rubicon fellowship (NWO, 019.161LW.017), NHMRC EL1 fellowship (GNT1178339), and The Kids? Cancer Project. S.A.W. was supported by a training grant from NIH (T32-GM071339). J.R.D. was supported by an Early Career Seed Grant from the Victorian Cancer Agency (VCA). S.J.H. was supported by a Postdoctoral Fellowship from the Cancer Council of Victoria (CCV). K.F.H. was supported by a NHMRC Senior Research Fellowship (APP1078220). J.W. and K.P. were supported by Academy of Finland (294850) and Finnish Cancer Foundation. K.P. was supported by Finnish Cultural Foundation. The VCFG ACRF Translational Reverse Phase Protein Array platform (K.J.S.) is funded by the Australian Cancer Research Foundation (ACRF), University of Melbourne Collaborative Research Infrastructure Program, and the PMCC Foundation. The PMCC Foundation and ACRF provide generous support for equipment and core facilities. S.J.V. S.A.W. J.R.D. D.A.K. E.B. I.T. C.J.K. M.C. S.B. Z.F. J.H.A.V. B.P.M. I.Y.K. S.J.H. M.J.K. E.D. M.Z. B.B. V.M. S.O. and A.N. performed experiments, data analysis, next-generation sequencing analysis, and data interpretation. J.J.S. performed mass spectroscopy. K.P. O.K. K.F.H. M.O. E.D.H. J.W. K.J.S. G.G. and N.G. provided critical reagents and advice. S.J.V. R.W.J. and A.G. contributed to data interpretation and study supervision. S.J.V. S.A.W. J.R.D. A.G. and R.W.J. wrote the manuscript, which was proofread and edited by all co-authors prior to submission. The Johnstone laboratory receives funding support from Roche, BMS, Astra Zeneca, and MecRx. R.W.J. is a shareholder in MecRx and receives consultancy payments. Funding Information: The Johnstone laboratory receives funding support from Roche, BMS, Astra Zeneca, and MecRx. R.W.J. is a shareholder in MecRx and receives consultancy payments. Publisher Copyright: © 2021 Elsevier Inc. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.
PY - 2021/6/10
Y1 - 2021/6/10
N2 - Gene expression by RNA polymerase II (RNAPII) is tightly controlled by cyclin-dependent kinases (CDKs) at discrete checkpoints during the transcription cycle. The pausing checkpoint following transcription initiation is primarily controlled by CDK9. We discovered that CDK9-mediated, RNAPII-driven transcription is functionally opposed by a protein phosphatase 2A (PP2A) complex that is recruited to transcription sites by the Integrator complex subunit INTS6. PP2A dynamically antagonizes phosphorylation of key CDK9 substrates including DSIF and RNAPII-CTD. Loss of INTS6 results in resistance to tumor cell death mediated by CDK9 inhibition, decreased turnover of CDK9 phospho-substrates, and amplification of acute oncogenic transcriptional responses. Pharmacological PP2A activation synergizes with CDK9 inhibition to kill both leukemic and solid tumor cells, providing therapeutic benefit in vivo. These data demonstrate that fine control of gene expression relies on the balance between kinase and phosphatase activity throughout the transcription cycle, a process dysregulated in cancer that can be exploited therapeutically.
AB - Gene expression by RNA polymerase II (RNAPII) is tightly controlled by cyclin-dependent kinases (CDKs) at discrete checkpoints during the transcription cycle. The pausing checkpoint following transcription initiation is primarily controlled by CDK9. We discovered that CDK9-mediated, RNAPII-driven transcription is functionally opposed by a protein phosphatase 2A (PP2A) complex that is recruited to transcription sites by the Integrator complex subunit INTS6. PP2A dynamically antagonizes phosphorylation of key CDK9 substrates including DSIF and RNAPII-CTD. Loss of INTS6 results in resistance to tumor cell death mediated by CDK9 inhibition, decreased turnover of CDK9 phospho-substrates, and amplification of acute oncogenic transcriptional responses. Pharmacological PP2A activation synergizes with CDK9 inhibition to kill both leukemic and solid tumor cells, providing therapeutic benefit in vivo. These data demonstrate that fine control of gene expression relies on the balance between kinase and phosphatase activity throughout the transcription cycle, a process dysregulated in cancer that can be exploited therapeutically.
KW - cancer
KW - CDK9
KW - CRISPR-Cas9 screen
KW - CTD
KW - Integrator
KW - pause-release
KW - phosphatase
KW - PP2A
KW - PP2A activation
KW - RNA polymerase II
KW - transcriptional elongation
UR - http://www.scopus.com/inward/record.url?scp=85106336505&partnerID=8YFLogxK
U2 - 10.1016/j.cell.2021.04.022
DO - 10.1016/j.cell.2021.04.022
M3 - Article
C2 - 34004147
AN - SCOPUS:85106336505
SN - 0092-8674
VL - 184
SP - 3143-3162.E32
JO - Cell
JF - Cell
IS - 12
ER -