title: Multivalent recognition of histone tails by the PHD fingers of CHD5.
abstract: The chromodomain, helicase, DNA-binding protein 5 (CHD5) is a chromatin remodeling enzyme which is implicated in tumor suppression. In this study, we demonstrate the ability of the CHD5 PHD fingers to specifically recognize the unmodified N-terminus of histone H3. We use two distinct modified peptide-library platforms (beads and glass slides) to determine the detailed histone binding preferences of PHD(1) and PHD(2) alone and the tandem PHD(1-2) construct. Both domains displayed similar binding preferences for histone H3, where modification (e.g., methylation, acetylation, and phosphorylation) at H3R2, H3K4, H3T3, H3T6, and H3S10 disrupts high-affinity binding, and the three most N-terminal amino acids (ART) are crucial for binding. The tandem CHD5-PHD(1-2) displayed similar preferences to those displayed by each PHD finger alone. Using NMR, surface plasmon resonance, and two novel biochemical assays, we demonstrate that CHD5-PHD(1-2) simultaneously engages two H3 N-termini and results in a 4-11-fold increase in affinity compared with either PHD finger alone. These studies provide biochemical evidence for the utility of tandem PHD fingers to recruit protein complexes at targeted genomic loci and provide the framework for understanding how multiple chromatin-binding modules function to interpret the combinatorial PTM capacity written in chromatin.
authors: Department of Biomolecular Chemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Samuel S
authors: SS
authors: Oliver
authors: Catherine A
authors: CA
authors: Musselman
authors: Rajini
authors: R
authors: Srinivasan
authors: John P
authors: JP
authors: Svaren
authors: Tatiana G
authors: TG
authors: Kutateladze
authors: John M
authors: JM
authors: Denu
mesh_terms: D000595:Amino Acid Sequence; D002843:Chromatin / Q000378:metabolism; D004265:DNA Helicases / Q000737:chemistry*; D004268:DNA-Binding Proteins / Q000737:chemistry; D006657:Histones / Q000737:chemistry*; D018398:Homeodomain Proteins / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D063146:Polycomb-Group Proteins; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D014157:Transcription Factors / Q000737:chemistry
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; D006657:Histones; D018398:Homeodomain Proteins; D009419:Nerve Tissue Proteins; C112309:PHF1 protein, human; C117905:PHF2 protein, human; D063146:Polycomb-Group Proteins; D014157:Transcription Factors; D004265:DNA Helicases; C471926:CHD5 protein, human