title: Inhibition of histone binding by supramolecular hosts.
abstract: The tandem PHD (plant homeodomain) fingers of the CHD4 (chromodomain helicase DNA-binding protein 4) ATPase are epigenetic readers that bind either unmodified histone H3 tails or H3K9me3 (histone H3 trimethylated at Lys⁹). This dual function is necessary for the transcriptional and chromatin remodelling activities of the NuRD (nucleosome remodelling and deacetylase) complex. In the present paper, we show that calixarene-based supramolecular hosts disrupt binding of the CHD4 PHD2 finger to H3K9me3, but do not affect the interaction of this protein with the H3K9me0 (unmodified histone H3) tail. A similar inhibitory effect, observed for the association of chromodomain of HP1γ (heterochromatin protein 1γ) with H3K9me3, points to a general mechanism of methyl-lysine caging by calixarenes and suggests a high potential for these compounds in biochemical applications. Immunofluorescence analysis reveals that the supramolecular agents induce changes in chromatin organization that are consistent with their binding to and disruption of H3K9me3 sites in living cells. The results of the present study suggest that the aromatic macrocyclic hosts can be used as a powerful new tool for characterizing methylation-driven epigenetic mechanisms.
authors.affiliation: *Department of Pharmacology, University of Colorado School of Medicine, Aurora, CO 80045, U.S.A.
authors.forename: Hillary F
authors.initials: HF
authors.lastname: Allen
authors.affiliation: †Department of Chemistry, University of Victoria, Victoria, BC, Canada, V8W 3V6.
authors.forename: Kevin D
authors.initials: KD
authors.lastname: Daze
authors.affiliation: ‡Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, U.S.A.
authors.forename: Takashi
authors.initials: T
authors.lastname: Shimbo
authors.affiliation: ‡Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, U.S.A.
authors.forename: Anne
authors.initials: A
authors.lastname: Lai
authors.affiliation: *Department of Pharmacology, University of Colorado School of Medicine, Aurora, CO 80045, U.S.A.
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Musselman
authors.affiliation: ‡Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, U.S.A.
authors.forename: Jennifer K
authors.initials: JK
authors.lastname: Sims
authors.affiliation: ‡Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, U.S.A.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Wade
authors.affiliation: †Department of Chemistry, University of Victoria, Victoria, BC, Canada, V8W 3V6.
authors.forename: Fraser
authors.initials: F
authors.lastname: Hof
authors.affiliation: *Department of Pharmacology, University of Colorado School of Medicine, Aurora, CO 80045, U.S.A.
authors.forename: Tatiana G
authors.initials: TG
authors.lastname: Kutateladze
mesh_terms: D001324:Autoantigens / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D047250:Calixarenes / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D042002:Chromatin Assembly and Disassembly / Q000187:drug effects*; D002868:Chromosomal Proteins, Non-Histone / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015195:Drug Design; D044127:Epigenesis, Genetic / Q000187:drug effects; D057809:HEK293 Cells; D006657:Histones / Q000037:antagonists & inhibitors* / Q000378:metabolism; D018398:Homeodomain Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007202:Indicators and Reagents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D008239:Lysine / Q000031:analogs & derivatives / Q000378:metabolism; D008745:Methylation; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D010446:Peptide Fragments / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D054730:Protein Interaction Domains and Motifs / Q000187:drug effects; D011499:Protein Processing, Post-Translational; D021122:Protein Subunits / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D001324:Autoantigens; C518294:CBX3 protein, human; C115323:CHD4 protein, human; D002868:Chromosomal Proteins, Non-Histone; D006657:Histones; D018398:Homeodomain Proteins; D007202:Indicators and Reagents; D010446:Peptide Fragments; D021122:Protein Subunits; D011994:Recombinant Proteins; D047250:Calixarenes; C003712:trimethyllysine; C490190:EGLN1 protein, human; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex; D008239:Lysine