title: Differentially Isotope-Labeled Nucleosomes To Study Asymmetric Histone Modification Crosstalk by Time-Resolved NMR Spectroscopy.
abstract: Post-translational modifications (PTMs) of histones regulate chromatin structure and function. Because nucleosomes contain two copies each of the four core histones, the establishment of different PTMs on individual "sister" histones in the same nucleosomal context, that is, asymmetric histone PTMs, are difficult to analyze. Here, we generated differentially isotope-labeled nucleosomes to study asymmetric histone modification crosstalk by time-resolved NMR spectroscopy. Specifically, we present mechanistic insights into nucleosomal histone H3 modification reactions in cis and in trans, that is, within individual H3 copies or between them. We validated our approach by using the H3S10phK14ac crosstalk mechanism, which is mediated by the Gcn5 acetyltransferase. Moreover, phosphorylation assays on methylated substrates showed that, under certain conditions, Haspin kinase is able to produce nucleosomes decorated asymmetrically with two distinct types of PTMs.
authors.affiliation: Department of Structural Biology, Leibniz-Institut für Molekulare Pharmakologie, Robert-Rössle-Strasse 10, 13125, Berlin, Germany. liokatis@fmp-berlin.de.
authors.forename: Stamatios
authors.identifier: 0000-0002-1940-9282
authors.initials: S
authors.lastname: Liokatis
authors.affiliation: Department of Chemical Biology, Leibniz-Institut für Molekulare Pharmakologie, Robert-Rössle-Strasse 10, 13125, Berlin, Germany.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Klingberg
authors.affiliation: Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, 16802-1014, USA.
authors.forename: Song
authors.initials: S
authors.lastname: Tan
authors.affiliation: Department of Chemical Biology, Leibniz-Institut für Molekulare Pharmakologie, Robert-Rössle-Strasse 10, 13125, Berlin, Germany.|Chemische Biologie, Interfakultäres Institut für Biochemie, Universität Tübingen, Hoppe-Seyler-Strasse 4, 72076, Tübingen, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Schwarzer
mesh_terms: D051548:Histone Acetyltransferases / Q000378:metabolism; D006657:Histones / Q000737:chemistry* / Q000378:metabolism; D007553:Isotope Labeling; D019906:Nuclear Magnetic Resonance, Biomolecular; D009707:Nucleosomes / Q000737:chemistry* / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D013997:Time Factors
chemical_list: D006657:Histones; D009707:Nucleosomes; D051548:Histone Acetyltransferases
keywords: NMR spectroscopy; chromatin biology; epigenetics; nucleosomes; protein modifications