title: An Assay for Measuring Histone Variant Exchange within Nucleosomes In Vitro.
abstract: The incorporation of histone variants into specific chromatin regions is a mechanism by which cells can regulate many important biological processes. One such example is H2A.Z, a highly conserved variant of H2A that is incorporated in genomic regulatory regions and contributes to control gene expression. H2A.Z variant exchange involves the removal of H2A-H2B dimers from a preassembled nucleosome and their replacement with H2A.Z-H2B dimers. A specific family of chromatin remodeling complexes, homologous to the yeast Swr1 complex, have been shown to be capable of this histone exchange activity both in vivo and in vitro. Here, we describe an assay to measure the histone H2A.Z exchange activity of recombinant human p400 on immobilized mononucleosomes in vitro. The assay can be adapted to other histone exchange complexes/catalytic subunits purified from any species.
authors.affiliation: Département de biologie, Faculté des sciences, Université de Sherbrooke, 2500 boul. de l'Université, Sherbrooke, QC, Canada, J1K 2R1.
authors.forename: Liette
authors.initials: L
authors.lastname: Laflamme
authors.affiliation: Department of Biology, Université de Sherbrooke, 2500 boul. de l'Université, Sherbrooke, QC, Canada, J1K 2R1.
authors.forename: Benoit
authors.initials: B
authors.lastname: Guillemette
authors.affiliation: Department of Biology, Université de Sherbrooke, 2500 boul. de l'Université, Sherbrooke, QC, Canada, J1K 2R1. luc.gaudreau@usherbrooke.ca.
authors.forename: Luc
authors.initials: L
authors.lastname: Gaudreau
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002843:Chromatin / Q000378:metabolism; D042002:Chromatin Assembly and Disassembly / Q000235:genetics / Q000502:physiology; D006367:HeLa Cells; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D009707:Nucleosomes / Q000378:metabolism*; D061987:Sf9 Cells
chemical_list: D002843:Chromatin; D006657:Histones; D009707:Nucleosomes
keywords: Chromatin; Exchange; H2A.Z; Histone variant; Nucleosome; Swr1; p400