First critical repressive H3K27me3 marks in embryonic stem cells identified using designed protein inhibitor
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- Univ. of Washington, Seattle, WA (United States)
- Dana Farber Cancer Inst., Boston, MA (United States); Boston Children’s Hospital, MA (United States); Harvard Medical School, Boston, MA (United States)
- Univ. of Toronto, ON (Canada)
- Univ. of Toronto, ON (Canada); Jinggangshan Univ., Jiangxi Province (People’s Republic of China)
- Univ. of Toronto, ON (Canada); Univ. of Science and Technology of China, Hefei (China)
- Univ. of Washington, Seattle, WA (United States); Howard Hughes Medical Inst., Seattle, WA (United States)
- Dana Farber Cancer Inst., Boston, MA (United States); Boston Children’s Hospital, MA (United States); Harvard Medical School, Boston, MA (United States); Howard Hughes Medical Inst., Boston, MA (United States)
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, in this work, 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.
- Research Organization:
- Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
- Sponsoring Organization:
- National Institutes of Health (NIH); NHLBI Progenitor Cell Biology Consortium; WRF Innovation Fellowship Program; University of Washington’s Proteomics Resource Grant; Defense Threat Reduction Agency (DTRA); Claudia Adams Barr Grant; National Institute of General Medical Sciences; USDOE Office of Science (SC); 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; Innovative Medicines Initiative; 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; Wellcome Trust
- Grant/Contract Number:
- R01GM097372; R01GM97372-03S1; R01GM083867; U01HL099997; U01HL099993; P01GM081619; UWPR95794; GM103533; P41 GM103403; AC02-06CH11357; OGI-055; 115766; 092809/Z/10/Z
- OSTI ID:
- 1430334
- Journal Information:
- Proceedings of the National Academy of Sciences of the United States of America, Vol. 114, Issue 38; ISSN 0027-8424
- Publisher:
- National Academy of SciencesCopyright Statement
- Country of Publication:
- United States
- Language:
- ENGLISH
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