Fig. 3: DNA sequence selectivity of MORFWHs.
From: MORF and MOZ acetyltransferases target unmethylated CpG islands through the winged helix ___domain
a Analysis of DNA binding selectivity of MORFWH2 in universal oligonucleotide PBMs. One of the replicates met significance criteria. b EMSAs of 15 bp A-rich, 16 bp unmethylated CpG, and 16 bp methylated CpG dsDNA in the presence of increasing amounts of GST-MORFWH2. DNA:protein ratio is shown below the gel images. c, d EMSA-derived binding curves used to determine Kd values for the interaction of MORFWH2 with the indicated DNA. Data represent average ± SD of three independent experiments. n = 3. e Analysis of DNA binding specificity of MORFWH1 in universal oligonucleotide PBMs. f EMSAs of 16 bp unmethylated CpG, 16 bp methylated CpG, and 15 bp A-rich dsDNA in the presence of increasing amounts of His-MORFWH1. DNA:protein ratio is shown below the gel images. g EMSA-derived binding curves used to determine Kd values for the interaction of His-MORFWH1 with 16 bp unmethylated CpG dsDNA. Data represent average ± SD of four independent experiments. n = 4. h Electrostatic surface potential of the alpha fold (AF) computational model of MORFWH1 from UniProt (Q8WYB5), with blue and red colors representing positive and negative charges, respectively. Two positively charged regions of MORFWH1 are indicated by green dotted ovals, with lysine and arginine residues labeled. i Superimposition of the AF model of MORFWH1 from UniProt (Q8WYB5) (blue) and the crystal structure of the DNA-bound WH from SAMD1 (salmon), with CpG colored green (PDB ID 6LUI). The mutated residues of MORFWH1 are shown as sticks and labeled. j, k EMSA of 147 bp 601 DNA in the presence of increasing amounts of K19A/K21A/K22A MORFWH1 (j) and K24A/R26A/K66A MORFWH1 (k) mutants. DNA:protein ratio is shown below the gel images. l–n Overlay of 1H,15N HSQC spectra of His-MORFWH1, wild type (l), K19A/K21A/K22A (m) or K24A/R26A/K66A (n) in the presence of increasing amount of CpG-DNA16. Spectra are color coded according to the protein:DNA molar ratio. Source data are provided as a Source Data file.
