title: Binding of the CHD4 PHD2 finger to histone H3 is modulated by covalent modifications.
abstract: CHD4 (chromodomain helicase DNA-binding protein 4) ATPase is a major subunit of the repressive NuRD (nucleosome remodelling and deacetylase) complex, which is involved in transcriptional regulation and development. CHD4 contains two PHD (plant homeodomain) fingers of unknown function. Here we show that the second PHD finger (PHD2) of CHD4 recognizes the N-terminus of histone H3 and that this interaction is facilitated by acetylation or methylation of Lys9 (H3K9ac and H3K9me respectively) but is inhibited by methylation of Lys4 (H3K4me) or acetylation of Ala1 (H3A1ac). An 18 microM binding affinity toward unmodified H3 rises to 0.6 microM for H3K9ac and to 0.9 microM for H3K9me3, whereas it drops to 2.0 mM for H3K4me3, as measured by tryptophan fluorescence and NMR. A peptide library screen further shows that phosphorylation of Thr3, Thr6 or Ser10 abolishes this interaction. A model of the PHD2-H3 complex, generated using a combination of NMR, data-driven docking and mutagenesis data, reveals an elongated site on the PHD2 surface where the H3 peptide is bound. Together our findings suggest that the PHD2 finger plays a role in targeting of the CHD4/NuRD complex to chromatin.
authors.affiliation: Department of Pharmacology, School of Medicine, University of Colorado Denver, Aurora, CO 80045, USA.
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Musselman
authors.forename: Robyn E
authors.initials: RE
authors.lastname: Mansfield
authors.forename: Adam L
authors.initials: AL
authors.lastname: Garske
authors.forename: Foteini
authors.initials: F
authors.lastname: Davrazou
authors.forename: Ann H
authors.initials: AH
authors.lastname: Kwan
authors.forename: Samuel S
authors.initials: SS
authors.lastname: Oliver
authors.forename: Heather
authors.initials: H
authors.lastname: O'Leary
authors.forename: John M
authors.initials: JM
authors.lastname: Denu
authors.forename: Joel P
authors.initials: JP
authors.lastname: Mackay
authors.forename: Tatiana G
authors.initials: TG
authors.lastname: Kutateladze
mesh_terms: D000107:Acetylation; D001324:Autoantigens / Q000737:chemistry* / Q000378:metabolism*; D001665:Binding Sites; D002843:Chromatin / Q000378:metabolism; D004265:DNA Helicases / Q000737:chemistry* / Q000378:metabolism*; D051548:Histone Acetyltransferases / Q000378:metabolism / Q000502:physiology*; D000076983:Histone Methyltransferases; D011495:Histone-Lysine N-Methyltransferase / Q000378:metabolism / Q000502:physiology*; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008745:Methylation; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex; D008954:Models, Biological; D011485:Protein Binding; D017434:Protein Structure, Tertiary / Q000502:physiology; D013379:Substrate Specificity
chemical_list: D001324:Autoantigens; C115323:CHD4 protein, human; D002843:Chromatin; D006657:Histones; D018398:Homeodomain Proteins; D000076983:Histone Methyltransferases; D011495:Histone-Lysine N-Methyltransferase; D051548:Histone Acetyltransferases; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex; D004265:DNA Helicases