Résumé
The polycomb repressive complex 2 (PRC2) histone methyltransferase plays a central role in epigenetic regulation in development and in cancer, and hence to interrogate its role in a specific developmental transition, methods are needed for disrupting function of the complex with high temporal and spatial precision. The catalytic and substrate recognition functions of PRC2 are coupled by binding of the N-terminal helix of the Ezh2 methylase to an extended groove on the EED trimethyl lysine binding subunit. Disrupting PRC2 function can in principle be achieved by blocking this single interaction, but there are few approaches for blocking specific protein–protein interactions in living cells and organisms. Here, we describe the computational design of proteins that bind to the EZH2 interaction site on EED with subnanomolar affinity in vitro and form tight and specific complexes with EED in living cells. Induction of the EED binding proteins abolishes H3K27 methylation in human embryonic stem cells (hESCs) and at all but the earliest stage blocks self-renewal, pinpointing the first critical repressive H3K27me3 marks in development.
| langue originale | Anglais |
|---|---|
| Pages (de - à) | 10125-10130 |
| Nombre de pages | 6 |
| journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 114 |
| Numéro de publication | 38 |
| Les DOIs | |
| Etat de la publication | Publié - 19 sept. 2017 |
| Modification externe | Oui |
Financement
ACKNOWLEDGMENTS. We thank Prof. Ware and members of the H.R.-B. laboratory for helpful discussions during this work and Jennifer Hesson for help with 5iLA hESC transition and culturing. This work is supported by NIH Grants R01GM097372, R01GM97372-03S1, and R01GM083867 and NHLBI Progenitor Cell Biology Consortium Grants U01HL099997 and U01HL099993 (to H.R.-B.); the WRF Innovation Fellowship Program (S.L. and Y.X.); NIH Grant P01GM081619 (to H.R.-B. and R.T.M.); University of Washington’s Proteomics Resource Grant UWPR95794 (NIH/NIGMS); the Defense Threat Reduction Agency (J.D.M. and D.B.); NIH Yeast Resource Center Grant GM103533 (to J.D.M., D.B., and L.T.D.); and a Claudia Adams Barr grant (to W.K.). This work is partly based upon research conducted at the North-eastern Collaborative Access Team beamlines, which are funded by the National Institute of General Medical Sciences from NIH Grant P41 GM103403. This research used resources of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract DE-AC02-06CH11357. The Structural Genomics Consortium is a registered charity (1097737) that receives funds from AbbVie, Bayer Pharma AG, Boehringer Ingelheim, Canada Foundation for Innovation, Eshelman Institute for Innovation, the Canadian Institutes of Health Research (CIHR), Genome Canada, Ontario Genomics Institute (Grant OGI-055), Innovative Medicines Initiative (EU/EFPIA) (ULTRA-DD Grant 115766), GlaxoSmithKline, Janssen, Merck & Co., Novartis Pharma AG, Lilly Canada, the Novartis Research Foundation, the Ontario Ministry of Economic Development and Innovation, Pfizer, São Paulo Research Foundation-FAPESP, Takeda, and Wellcome Trust (Grant 092809/Z/10/Z).
| Bailleurs de fonds | Numéro du bailleur de fonds |
|---|---|
| National Heart, Lung, and Blood Institute | U01HL099997, U01HL099993 |
| National Institute of General Medical Sciences | R01GM097372, P01GM081619, T32GM007266, P41GM103533, P41GM103403, R01GM083867 |
| National Institute of Diabetes and Digestive and Kidney Diseases | R01DK103667, P30DK017047 |
| European commission | 115766 |
SDG des Nations Unies
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