Figure 1: Hm-DNA facilities histione tails recognition by full-length UHRF1.
From: Hemi-methylated DNA opens a closed conformation of UHRF1 to facilitate its histone recognition
(a) Colour-coded domain structure of human UHRF1. The boundaries of the domains are indicated with the numbers representing the amino-acid positions. Note that the conserved motif (green background) of the Linker (residues 286–306) and the Spacer (residues 587–674) bind to the TTD in a similar manner (Fig. 3b). (b) Hm-DNA facilities histone H3 and H3K9me3 recognition by UHRF1. Purified full-length UHRF1 was incubated with biotinylated H3 (1–21) or H3K9me3 (1–21) peptides in the presence or absence of hm-DNA (molar ratio UHRF1/hm-DNA=1:2). The mixture was immobilized onto streptavidin Sepharose beads. The bound proteins were analysed in SDS–PAGE followed by Coomassie blue staining. Sequences of the peptides are indicated in Supplementary Table 1. (c) Histone peptides do not affect hm-DNA-binding affinity of UHRF1. Full-length UHRF1 was incubated with biotinylated hm-DNA in the presence or absence of H3 (1–17) or H3K9me3 (1–17) peptides and analysed as in b. (d,e) Superimposed ITC enthalpy plots for binding of H3K9me3 peptide (1–17) to TTD–PHD and full-length UHRF1 (d), and H3 peptide (1–17) to the PHD and full-length UHRF1 (e). The estimated binding affinities (KD) are listed. The samples in the syringe (designated Sy hereafter) and cell are indicated.
