title: Chromatibody, a novel non-invasive molecular tool to explore and manipulate chromatin in living cells.
abstract: Chromatin function is involved in many cellular processes, its visualization or modification being essential in many developmental or cellular studies. Here, we present the characterization of chromatibody, a chromatin-binding single-domain, and explore its use in living cells. This non-intercalating tool specifically binds the heterodimer of H2A-H2B histones and displays a versatile reactivity, specifically labeling chromatin from yeast to mammals. We show that this genetically encoded probe, when fused to fluorescent proteins, allows non-invasive real-time chromatin imaging. Chromatibody is a dynamic chromatin probe that can be modulated. Finally, chromatibody is an efficient tool to target an enzymatic activity to the nucleosome, such as the DNA damage-dependent H2A ubiquitylation, which can modify this epigenetic mark at the scale of the genome and result in DNA damage signaling and repair defects. Taken together, these results identify chromatibody as a universal non-invasive tool for either in vivo chromatin imaging or to manipulate the chromatin landscape.
authors.affiliation: Toxalim, Université de Toulouse, INRA, Université de Toulouse 3 Paul Sabatier, 31027 Toulouse, France ITAV, Université de Toulouse, CNRS, UPS, 31106 Toulouse, France.
authors.forename: Denis
authors.initials: D
authors.lastname: Jullien
authors.affiliation: Toxalim, Université de Toulouse, INRA, Université de Toulouse 3 Paul Sabatier, 31027 Toulouse, France.
authors.forename: Julien
authors.initials: J
authors.lastname: Vignard
authors.affiliation: Toxalim, Université de Toulouse, INRA, Université de Toulouse 3 Paul Sabatier, 31027 Toulouse, France.
authors.forename: Yoann
authors.initials: Y
authors.lastname: Fedor
authors.affiliation: CRCT-UMR1037, Université de Toulouse, INSERM, 31037 Toulouse, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Béry
authors.affiliation: CRCT-UMR1037, Université de Toulouse, INSERM, 31037 Toulouse, France.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Olichon
authors.affiliation: Centre de Biologie du Développement UMR5547, 31062 Toulouse, France.
authors.forename: Michèle
authors.initials: M
authors.lastname: Crozatier
authors.affiliation: IPBS-UMR5089, Université de Toulouse, CNRS, 31077 Toulouse, France.
authors.forename: Monique
authors.initials: M
authors.lastname: Erard
authors.affiliation: IHAP, Université de Toulouse, INRA, ENVT, 31076 Toulouse, France.
authors.forename: Hervé
authors.initials: H
authors.lastname: Cassard
authors.affiliation: ITAV, Université de Toulouse, CNRS, UPS, 31106 Toulouse, France CHU de Toulouse, 31106 Toulouse, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Ducommun
authors.affiliation: Toxalim, Université de Toulouse, INRA, Université de Toulouse 3 Paul Sabatier, 31027 Toulouse, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Salles
authors.affiliation: Toxalim, Université de Toulouse, INRA, Université de Toulouse 3 Paul Sabatier, 31027 Toulouse, France gladys.mirey@toulouse.inra.fr.
authors.forename: Gladys
authors.identifier: 0000-0002-0095-1864
authors.initials: G
authors.lastname: Mirey
mesh_terms: D000818:Animals; D002161:Camelids, New World; D002843:Chromatin / Q000235:genetics* / Q000302:isolation & purification; D004249:DNA Damage / Q000235:genetics*; D006657:Histones / Q000378:metabolism; D009707:Nucleosomes / Q000235:genetics*; D054875:Ubiquitination / Q000235:genetics
chemical_list: D002843:Chromatin; D006657:Histones; D009707:Nucleosomes
keywords: Chromatin; Chromatin function; Epigenetic; Real-time imaging; Single-domain antibody