A chemical probe selectively inhibits G9a and GLP methyltransferase activity in cells (original) (raw)

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Acknowledgements

We thank A. Tumber for JMJD2E assay support; J. Moffat (University of Toronto) for the gift of shRNAs; R. Bristow (University Health Network) for RV221 and PC3 cells; T. Hajian and F. Syeda for protein purification; G. Senisterra for contributing to DSF and DSLS data analysis; M. Herold for graphical design and illustration; I. Korboukh, M. Herold and J. Yost for critical reading of the manuscript; and R. Trump and C. Yates for helpful discussion. The research described here was supported by the National Institute of General Medical Sciences, US National Institutes of Health (NIH; grant RC1GM090732), the Carolina Partnership and University Cancer Research Fund from the University of North Carolina at Chapel Hill, the US National Science Foundation (NSF), the Ontario Research Fund, the Ontario Ministry of Health and Long-term Care and the Structural Genomics Consortium. The Structural Genomics Consortium is a registered charity (number 1097737) that receives funds from the Canadian Institutes for Health Research (CIHR), the Canada Foundation for Innovation, Genome Canada through the Ontario Genomics Institute, GlaxoSmithKline, Karolinska Institutet, the Knut and Alice Wallenberg Foundation, the Ontario Innovation Trust, the Ontario Ministry for Research and Innovation, Merck & Co. Inc., the Novartis Research Foundation, the Swedish Agency for Innovation Systems, the Swedish Foundation for Strategic Research and the Wellcome Trust. C.H.A. holds a Canada Research Chair in Structural Genomics. V.L. is supported by a CIHR fellowship. A.P. is supported by grants from the CIHR (199170 and 186007) and from the NIH (MH074127, MH088413, DP3DK085698 and HG004535). A.P. is Tapscott Chair in Schizophrenia Studies and a Senior Fellow of the Ontario Mental Health Foundation. J.E. is supported by CIHR grant IG1-102956. B.A.G. is supported by grants from the NSF (Early Faculty CAREER award and CBET-0941143), the American Society for Mass Spectrometry and the NIH Office of the Director (DP2OD007447). P.A.D. is supported by NIH postdoctoral fellowship F32 NRSA.

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Author notes

  1. Masoud Vedadi, Dalia Barsyte-Lovejoy and Feng Liu: These authors contributed equally to this work.

Authors and Affiliations

  1. Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada
    Masoud Vedadi, Dalia Barsyte-Lovejoy, Abdellah Allali-Hassani, Gregory A Wasney, Alena Siarheyeva, Aiping Dong, Wolfram Tempel, Irene Chau, Aled Edwards, Peter J Brown & Cheryl H Arrowsmith
  2. Division of Medicinal Chemistry and Natural Products, Center for Integrative Chemical Biology and Drug Discovery, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA
    Feng Liu, Tim J Wigle, Xin Chen, Catherine D Simpson, Samantha G Pattenden, Jacqueline L Norris, Dmitri B Kireev, William P Janzen, Stephen V Frye & Jian Jin
  3. Developmental and Stem Cell Biology Program, SickKids Hospital, Toronto, Ontario, Canada
    Sylvie Rival-Gervier & James Ellis
  4. Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada
    James Ellis
  5. INRA, UMR 1198 Biologie du Développement et Reproduction, Jouy en Josas, France
    Sylvie Rival-Gervier
  6. Krembil Family Epigenetic Laboratory, Centre for Addiction and Mental Health, Toronto, Ontario, Canada
    Viviane Labrie, Sun-Chong Wang & Arturas Petronis
  7. Department of Chemistry, Princeton University, Princeton, New Jersey, USA
    Peter A DiMaggio & Benjamin A Garcia
  8. Institute of Systems Biology and Bioinformatics, National Central University, Jhongli City, Taiwan
    Sun-Chong Wang
  9. National Institute of Mental Health Psychoactive Drug Screening Program, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA
    Thomas J Mangano, Xi-ping Huang & Bryan L Roth
  10. Department of Biochemistry and Biophysics, UNC Macromolecular Interactions Facility, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA
    Ashutosh Tripathy
  11. Campbell Family Cancer Research Institute and Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, Toronto, Ontario, Canada
    Cheryl H Arrowsmith

Authors

  1. Masoud Vedadi
  2. Dalia Barsyte-Lovejoy
  3. Feng Liu
  4. Sylvie Rival-Gervier
  5. Abdellah Allali-Hassani
  6. Viviane Labrie
  7. Tim J Wigle
  8. Peter A DiMaggio
  9. Gregory A Wasney
  10. Alena Siarheyeva
  11. Aiping Dong
  12. Wolfram Tempel
  13. Sun-Chong Wang
  14. Xin Chen
  15. Irene Chau
  16. Thomas J Mangano
  17. Xi-ping Huang
  18. Catherine D Simpson
  19. Samantha G Pattenden
  20. Jacqueline L Norris
  21. Dmitri B Kireev
  22. Ashutosh Tripathy
  23. Aled Edwards
  24. Bryan L Roth
  25. William P Janzen
  26. Benjamin A Garcia
  27. Arturas Petronis
  28. James Ellis
  29. Peter J Brown
  30. Stephen V Frye
  31. Cheryl H Arrowsmith
  32. Jian Jin

Contributions

M.V., A.A.-H., A.S. and I.C. performed SAHH-coupled, fluorescence-polarization, DSF, DSLS and DNMT1 assays; D.B.-L. developed and performed in-cell western, MTT, ChIP, gene expression, clonogenicity, western blotting and immunofluorescence studies; F.L. developed the synthetic route to UNC0638 and UNC0737 and synthesized the compounds; S.R.-G. and J.E. developed and performed mES cell studies; V.L., S.-C.W. and A.P. performed H3K9me2 genomic localization and DNA methylation analysis; T.J.W. and W.P.J. performed mechanism-of-action studies; P.A.D. and B.A.G. performed MS-based proteomics studies; M.V., G.A.W., A.D., W.T., D.B.K. and C.H.A. solved and analyzed the X-ray crystal structure of the G9a-UNC0638-SAH complex; T.J.W. and A.T. performed SPR studies; X.C. and S.G.P. performed UNC0638 stability studies; S.G.P. performed RT-qPCR studies; T.J.M., X.-p.H. and B.L.R. performed GPCR selectivity studies; C.D.S. and W.P.J. performed kinase selectivity studies; J.L.N. purified proteins; J.J. designed UNC0638 and UNC0737; C.H.A., J.J., S.V.F., M.V., D.B.-L., P.J.B., J.E., S.R.-G., A.P., V.L., B.A.G., T.J.W. and A.E. designed studies and discussed results; J.J., C.H.A., D.B.-L., S.R.-G., M.V., V.L., S.V.F., J.E., A.P., B.A.G., P.J.B. and T.J.W. wrote the paper.

Corresponding authors

Correspondence toCheryl H Arrowsmith or Jian Jin.

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The authors declare no competing financial interests.

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Vedadi, M., Barsyte-Lovejoy, D., Liu, F. et al. A chemical probe selectively inhibits G9a and GLP methyltransferase activity in cells.Nat Chem Biol 7, 566–574 (2011). https://doi.org/10.1038/nchembio.599

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