Figure 5: The Spacer facilitates hm-DNA–SRA interaction and DNMT1–UHRF1 interaction.
From: Hemi-methylated DNA opens a closed conformation of UHRF1 to facilitate its histone recognition
(a) Superimposed ITC enthalpy plots for hm-DNA-binding affinities of the SRA, the Spacer and SRA–Spacer. (b,c) Superimposed fluorescence polarization (FP) plots for hm-DNA-binding affinities of truncations or full-length UHRF1. The estimated binding affinities (KD) are listed above. (d) Subcellular localization of GFP-tagged wild-type or indicated mutants of UHRF1 in NIH3T3 cells. The percentages of cells showing co-localization with DAPI foci were counted from at least 100 cells and shown on the left of the corresponding representative confocal microscopy. The experiment was repeated three times in the same condition. Scale bar, 5 μm. (e) GST pull-down experiment for the interactions between wild-type or truncations of UHRF1 and RFTSDNMT1 as described in Fig. 2a. (f) Working model for hm-DNA-mediated conformational changes of UHRF1, as described in the Discussion. ND, not determined; Sy., syringe.
