Fig. 5. The C397R mutation boosts DRM2-mediated CHG methylation.
(A) In vitro methylation assay of DRM2 or C397R on DNA with different sequence contexts. CG, (GAC)12; CTA, (TAC)12; CAA, (AAC)12; CTG, (TGC)12. Data are means ± SD (n = 3 replicates). Statistical analysis used two-tailed Student’s t test for the difference from WT: *P < 0.05, ***P < 0.001, and ****P < 0.0001. (B) Ribbon representation of DRM2C397R bound to CCG DNA and SAH. Hydrogen-bonding interactions formed between the side chain of R397 and G12 are depicted as dashed lines in expanded view. The bases of C11 and G12 in the expanded view are colored yellow. (C) Metaplots showing average methylation level of DRM2, C397R, and ddc over TEs for CG, CHG, and CHH contexts. (D) Representative genomic regions of two TEs on chromosome 3 (AT3TE666360 and AT3TE28430/AT3TE28440) showing the methylation levels of Col-0, ddc, DRM2/ddc, and C397R/ddc. (E) Bar chart showing the total number of DMCs in each context of DRM2/ddc and C397R/ddc called against ddc. (F) Motif of the 4 nucleotides upstream and 5 nucleotides downstream of hyper DMCs in C397R called against ddc (n = 29,347).