1. Volume 26, Issue 2, Pages 304-311.e3 (February 2018)
An Intramolecular Interaction of UHRF1 Reveals Dual Control for Its Histone Association 
Linfeng Gao, Xiao-Feng Tan, Shen Zhang, Tianchen Wu, Zhi-Min Zhang, Hui-wang Ai, Jikui Song 
Structure 
Volume 26, Issue 2, Pages e3 (February 2018)
DOI: /j.str
Copyright © 2017 Elsevier Ltd Terms and Conditions

2. Structure 2018 26, 304-311.e3DOI: (10.1016/j.str.2017.12.016)
Copyright © 2017 Elsevier Ltd Terms and Conditions

3. Figure 1 Crystal Structure of the UHRF1 TTD-PBR Complex
(A) Domain architecture of human UHRF1, with individual domains labeled with residue numbers. The TTD domain of zebrafish UHRF1 (zUHRF1TTD) used for structure determination is also shown.
(B) Ribbon representation of zUHRF1TTD (cyan) bound to the PBR peptide (yellow sticks).
(C) Surface representation of the zUHRF1TTD-PBR complex.
(D) Close-up view of the intermolecular interaction between zUHRF1TTD and PBR. The water molecules are shown in sphere representation. The hydrogen bonds are shown as dashed lines.
(E) Schematic representation of the zUHRF1TTD-PBR interaction. The residues from zUHRF1TTD and PBR are colored in magenta and black, respectively. Yellow: hydrophobic contact.
See also Figure S1.
Structure  , e3DOI: ( /j.str )
Copyright © 2017 Elsevier Ltd Terms and Conditions

4. Figure 2 ITC Binding Assays for Human UHRF1 TTD and PBR
(A) Mutational effects of the UHRF1 TTD domain on the TTD-PBR interaction.
(B) Mutational effects of the UHRF1 PBR peptide on the TTD-PBR interaction.
See also Table S1.
Structure  , e3DOI: ( /j.str )
Copyright © 2017 Elsevier Ltd Terms and Conditions

5. Figure 3 UHRF1 PBR and Histone H3K9me3 Compete on TTD Binding
(A) Structural comparison of the UHRF1 TTD-PBR complex with the UHRF1 TTD-PHD-H3K9me3 complex (PDB: 4GY5), with individual domains and peptides labeled. The zinc ions are shown as purple spheres.
(B) Structural overlay of the TTD-bound PBR peptide (yellow) and the TTD-PHD linker (magenta), with the aligned residues shown in stick representation.
(C) The side chain of H3K9me3 inserted into the aromatic cage formed by residues F152, Y188, and Y191 of human UHRF1 TTD (PDB: 4GY5).
(D) The side chain of P656 embedded in the equivalent aromatic cage formed by residues F154, Y180, and Y183 of zUHRF1TTD.
(E) ITC binding curves of the PBR peptide over the TTD domain or the TTD-PHD dual domains of human UHRF1, in the absence or presence of unmodified H31-22 or H31-22K9me3 peptides. The average and SD of the dissociation constants were derived from two independent measurements. ND, not determinable due to undetectable or non-stoichiometric binding.
See also Figures S1 and S2 and Table S1.
Structure  , e3DOI: ( /j.str )
Copyright © 2017 Elsevier Ltd Terms and Conditions

6. Figure 4
Coupling between the Conformational Transition of UHRF1 and Its Chromatin or USP7 Binding
(A) FRET ratio, EFRET = Iaccetpor/(Iaccetpor + Idonor), of CFP-UHRF1-YFP as function of the concentrations of hemimethylated DNA (HmDNA), USP7 UBL domains (USP7), H31-22K9me3 (H3K9me3) and H31-22 (H3) peptides. Plotted are the average and SD of the FRET ratio derived from two independent measurements.
(B) Pull-down assays of wild-type (WT) or R649A/P656G-mutated UHRF1 (residues 126–793) with H3K9me3 or unmodified H3 peptides. The gel image cropped from a full gel is boxed.
(C) A model for the conformational transition of UHRF1. (i) UHRF1 in free state is dominated by a “closed” conformation. (ii) Association of UHRF1 with hemimethylated DNA (HmDNA) and H3K9me3-modified nucleosome (H3K9me3-NCP) transits UHRF1 into an “open” conformation, permitting strong chromatin association and enhanced H3 ubiquitylation activity. (iii) Ubiquitylated histone H3 subsequently recruits DNMT1 for maintenance DNA methylation.
See also Figures S3–S5.
Structure  , e3DOI: ( /j.str )
Copyright © 2017 Elsevier Ltd Terms and Conditions