Recognition of Histone H3K14 Acylation by MORF Brianna J. Klein, Johayra Simithy, Xiaolu Wang, JaeWoo Ahn, Forest H. Andrews, Yi Zhang, Jacques Côté, Xiaobing Shi, Benjamin A. Garcia, Tatiana G. Kutateladze  Structure  Volume 25, Issue 4, Pages 650-654.e2 (April 2017) DOI: 10.1016/j.str.2017.02.003 Copyright © 2017 Elsevier Ltd Terms and Conditions

Structure 2017 25, 650-654.e2DOI: (10.1016/j.str.2017.02.003) Copyright © 2017 Elsevier Ltd Terms and Conditions

Figure 1 Acetyllysine Selectivity of the MORF DPF Domain (A) Binding affinities of the MORF DPF for indicated peptides were measured by tryptophan fluorescence. Values represent the average of three separate experiments. Error bars denote SD. (B) Representative binding curves used to determine KD. (C) Superimposed 1H,15N HSQC spectra of 15N-labeled MORF DPF, collected as indicated. H3K14acyl peptides were added stepwise. The intermediate exchange regime is commonly observed for binding of readers to histone peptides, indicating strong interaction. Spectra are color coded according to the protein/peptide molar ratio (inset). Structure 2017 25, 650-654.e2DOI: (10.1016/j.str.2017.02.003) Copyright © 2017 Elsevier Ltd Terms and Conditions

Figure 2 Structural Mechanism for the Recognition of H3K14bu by MORF (A) The crystal structure of the MORF DPF-H3K14bu complex. The DPF domain is depicted in ribbon diagram with the first and second PHD fingers colored yellow and gray, respectively, and H3K14bu peptide is colored green. The histone peptide residues and the residues of DPF involved in the interaction are labeled. Dashed lines indicate hydrogen bonds. (B) The electrostatic surface potential of the MORF DPF is shown using blue and red colors for positive and negative charges, respectively. (C) A zoom-in view of the K14bu-binding channel. Structure 2017 25, 650-654.e2DOI: (10.1016/j.str.2017.02.003) Copyright © 2017 Elsevier Ltd Terms and Conditions

Figure 3 Identification of Histone H3K14acyl PTMs In Vitro and In Vivo (A) In vitro H3K14 acylation profiles. Values represent the average of the relative abundances of enzymatically acylated H3K14 by six HATs (see C). Lysine modification abbreviations are: acetyl (ac), propionyl (pr), butyryl (bu), crotonyl (cr), malonyl (mal), succinyl (suc), β-hydroxybutyryl (bhb), and glutaryl (glu). (B) Bar plots showing the quantification of acylated H3K14 in HeLa cells. All results are shown as the average of three biological replicates; error bars denote SD. (C) In vitro specificity of HATs for H3K14acyl. Radar chart showing the H3K14 acylation activity of indicated HATs in the presence of acetyl-, propionyl-, crotonyl-, butyryl-, malonyl-, β-hydroxybutyryl-, succinyl-, and glutaryl-CoA. Each spoke represents a different HAT and the anchor represents the relative abundance of modified H3 peptide KSTGGKAPR (aa 9–17) at position H3K14. HATs Tip60 and MOF were only evaluated in the presence of acetyl- and butyryl-CoA. All results are shown as the average of three biological replicates. Structure 2017 25, 650-654.e2DOI: (10.1016/j.str.2017.02.003) Copyright © 2017 Elsevier Ltd Terms and Conditions