RESEARCH ARTICLE Conserved RNA-binding specificity of polycomb repressive complex 2 is achieved by dispersed amino acid patches in EZH2 Yicheng Long1, Ben Bolanos2†, Lihu Gong3,4†, Wei Liu2, Karen J Goodrich1, Xin Yang3,4, Siming Chen3,4, Anne R Gooding1, Karen A Maegley5, Ketan S Gajiwala2, Alexei Brooun2*, Thomas R Cech1*, Xin Liu3,4* 1Department of Chemistry and Biochemistry, BioFrontiers Institute, Howard Hughes Medical Institute, University of Colorado Boulder, Boulder, United States; 2Worldwide Medicinal Chemistry, Worldwide Research and Development, Pfizer Inc., San Diego, United States; 3Cecil H. and Ida Green Center for Reproductive Biology Sciences, Division of Basic Research, Department of Obstetrics and Gynecology, UT Southwestern Medical Center, Dallas, United States; 4Department of Biophysics, UT Southwestern Medical Center, Dallas, United States; 5Oncology Research Unit, Worldwide Research and Development, Pfizer Inc., San Diego, United States Abstract Polycomb repressive complex 2 (PRC2) is a key chromatin modifier responsible for methylation of lysine 27 in histone H3. PRC2 has been shown to interact with thousands of RNA *For correspondence: species in vivo, but understanding the physiological function of RNA binding has been hampered
[email protected] (AB); by the lack of separation-of-function mutants. Here, we use comprehensive mutagenesis and
[email protected] hydrogen deuterium exchange mass spectrometry (HDX-MS) to identify critical residues for RNA (TRC); interaction in PRC2 core complexes from Homo sapiens and Chaetomium thermophilum, for which
[email protected] (XL) crystal structures are known. Preferential binding of G-quadruplex RNA is conserved, surprisingly † These authors contributed using different protein elements. Key RNA-binding residues are spread out along the surface of equally to this work EZH2, with other subunits including EED also contributing, and missense mutations of some of Competing interest: See these residues have been found in cancer patients.