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. Author manuscript; available in PMC: 2020 Sep 25.
Published in final edited form as: J Am Chem Soc. 2019 Sep 17;141(38):15029–15039. doi: 10.1021/jacs.9b02321

Figure 5.

Figure 5.

Semisynthesis and activity of modified tJARID2 proteins. (a) Schematic of the two-piece ligation strategy used to prepare tJARID2 K116me2, tJARID2 K116me3, tJARID2 pS120, tJARID2 pS124, and tJARID2 pS126. For experimental details, see Supporting Information. (b) Schematic of the three-piece ligation strategy used to prepare tJARID2 K116me3/pS120, tJARID2 K116me3/pS124, and tJARID2 K116me3/ pS126. For experimental details, see Supporting Information. (c) Coomassie stain of SDS-PAGE analysis of all semisynthetic and recombinant tJARID2 constructs used in this study. (d) Relative activity of either PRC2 core complex or PRC2 core complex with an equimolar amount of the indicated tJARID2 construct on an unmodified 12mer nucleosome array substrate (Ar-3). Data from radio-HMT assays are plotted relative PRC2 core on Ar-3. Errors = s.e.m. (n = 3−4). (e) Relative activity of PRC2 with tJARID2 K116me3 or PRC2 with tJARID2 K116me3/pS120 on Ar-1 (containing H3K27me3 stimulating nucleosomes) or Ar-2 (containing H3K27R nonsubstrate nucleosomes). Data from radio-HMT assays are plotted relative PRC2- tJARID2 on Ar-2.