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. 2017 Nov 29;6:e31558. doi: 10.7554/eLife.31558

Figure 2. The minimal RNA-binding complex of ctPRC2 includes EED and EBD-BAM domains of EZH2, and additional affinity might be provided by the CXC-SET domains.

Figure 2.

(A) Purified ctPRC2 comprises two polypeptides: a 107 kDa peptide consisting of EZH2 fused with the VEFS domain of SUZ12, and a 67 kDa EED subunit. Individual domains are color coded as represented in structures of panel (B). (B) Purified ctPRC2 wild type was Coomassie stained on a SDS-PAGE gel (left). Sub-complexes of ctPRC2 were purified and tested for binding with (GGAA)10 RNA (right). Regulatory moiety contains EED and N-terminal half of EZH2 (ending at SANT1L domain), and catalytic moiety contains C-terminal half of EZH2 (starting at MCSS domain) fused with VEFS domain of SUZ12. Kdapp values and errors are mean and standard derivation of at least three binding experiments performed on different days. Minimal RNA-binding complex maintains the specificity of binding to G-quadruplex RNA. Regulatory moiety (reg) and minimal RNA-binding complex (mini; EED + EBD BAM of Ezh2) of ctPRC2. G-quadruplex-forming (GGAA)10 RNA and control (GA)20 RNA were tested for binding with the two complexes.