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. 2026 Feb 5;54(4):gkag089. doi: 10.1093/nar/gkag089

Figure 4.

Figure 4.

DNMT1 N-terminal mutants have decreased DNMT1 dynamics. (A) IUPred3 of DNMT1 showing the disordered domain (residues 100-400) and α-helical domain (residues 22-91). (B) N-terminal sequence of DNMT1, all Asp (A) and Glu (E) residues are highlighted in red, Pro (P) and Tyr (Y) are highlighted in yellow. Only the first 400 amino acids are shown. (C) Schematic of the DNMT1 N-terminal mutants, all D and E residues in the hatched region were mutated to alanine. Only the first 600 amino acids are shown. (D–G) Wildtype DNMT1 and N-terminal mutants were transfected into U2OS cells, and 18-24 h later live cell single molecule imaging was performed. Spot-on analysis showing the dynamics of DNMT1-Halo molecules. (D) Pie charts showing the distribution of fast or slow diffusing and chromatin-bound DNMT1 of various N-terminal mutants. (E–G) Violin plots showing the distribution of DNMT1-Halo in 17 to 214 cells across 2–5 replicates. All N-terminal DNMT1 mutants are less mobile compared to WT DNMT1. (G) The fraction bound of D/E-1-400 mutant is significantly increased compared to DNMT1 and other mutants. Student t-test, two tailed, **** P < 0.0001, ** P < 0.01.