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. Author manuscript; available in PMC: 2026 Jun 16.
Published in final edited form as: Mol Cell. 2026 Feb 24;86(5):834–850.e9. doi: 10.1016/j.molcel.2026.01.030

Figure 3: Linker length dependence of DNMT3A2/3B3 di-nucleosome bridging.

Figure 3:

a-c Gaussian filtered cryo-em maps of DNMT3A2/3B3 bound to the 10-5-10 (a) 10-6-10 (b) and 10-8-10 (c) di-nucleosomes. d Superimposed consensus cryo-EM maps from panel a-c. e Consensus cryo-EM maps of the 10-8-10 DNMT3A2/3B3 complex superimposed with the 10-5-10 (left) and 10-6-10 (right) DNMT3A2/3B3 complexes. Rotations in the lower nucleosome are indicated with the black angle indicator. Movement of the lower DNMT tetramer is indicated with an arrow. f Consensus Cryo-EM map of the 10-25-10 DNMT3A2/3B3 complex with missing density for the lower nucleosome. g Schematic of the experimental setup for binding. h A representative kymograph showing 3A2/3B3-GFP tetramer binding on the 8-bp linker di-nucleosome. i Schematic of the experimental setup for force manipulation. j Representative force–distance curve of a bare 8bp linker di-nucleosome-containing DNA tether showing force-induced transitions. The insert shows a zoom-in view of two example transitions for which the distance change (ΔL) and the transition force are recorded. k Fraction of trajectories on bare 8 or 25 bp linker di-nucleosome (N = 22 for 8bp linker, N=8 for 25bp linker) or 3A2/3B3-GFP tetramer bound 8 or 25 bp linker di-nucleosome (N = 12 for 8bp linker, N=8 for 25bp linker). l Representative force–distance curve of a 3A2/3B3-bound 8bp linker di-nucleosome as shown in j. m Low-force regime transition forces recorded from force-distance curves of bare (N = 10 from 10 independent tethers) or DNMT3A2/3B3-bound (N = 9 from 9 independent tethers) 8-bp di-nucleosomal DNA tethers. Significance was calculated using a two-tailed unpaired t-test (*P ≤ 0.05). See also Figure S5.