title: Insulators recruit histone methyltransferase dMes4 to regulate chromatin of flanking genes.
abstract: Chromosomal domains in Drosophila are marked by the insulator-binding proteins (IBPs) dCTCF/Beaf32 and cofactors that participate in regulating long-range interactions. Chromosomal borders are further enriched in specific histone modifications, yet the role of histone modifiers and nucleosome dynamics in this context remains largely unknown. Here, we show that IBP depletion impairs nucleosome dynamics specifically at the promoters and coding sequence of genes flanked by IBP binding sites. Biochemical purification identifies the H3K36 histone methyltransferase NSD/dMes-4 as a novel IBP cofactor, which specifically co-regulates the chromatin accessibility of hundreds of genes flanked by dCTCF/Beaf32. NSD/dMes-4 presets chromatin before the recruitment of transcriptional activators including DREF that triggers Set2/Hypb-dependent H3K36 trimethylation, nucleosome positioning, and RNA splicing. Our results unveil a model for how IBPs regulate nucleosome dynamics and gene expression through NSD/dMes-4, which may regulate H3K27me3 spreading. Our data uncover how IBPs dynamically regulate chromatin organization depending on distinct cofactors.
authors.affiliation: Laboratoire de Biologie Moléculaire Eucaryote (LBME), CNRS Université de Toulouse (UPS), Toulouse, France.
authors.forename: Priscillia
authors.initials: P
authors.lastname: Lhoumaud
authors.affiliation: Laboratoire de Biologie Moléculaire Eucaryote (LBME), CNRS Université de Toulouse (UPS), Toulouse, France.
authors.forename: Magali
authors.initials: M
authors.lastname: Hennion
authors.affiliation: Laboratoire de Biologie Moléculaire Eucaryote (LBME), CNRS Université de Toulouse (UPS), Toulouse, France.
authors.forename: Adrien
authors.initials: A
authors.lastname: Gamot
authors.affiliation: Systems Biology Center, National Heart, Lung and Blood Institute National Institutes of Health (NIH), Bethesda, MD, USA.
authors.forename: Suresh
authors.initials: S
authors.lastname: Cuddapah
authors.affiliation: Laboratoire de Biologie Moléculaire Eucaryote (LBME), CNRS Université de Toulouse (UPS), Toulouse, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Queille
authors.affiliation: Laboratoire de Biologie Moléculaire Eucaryote (LBME), CNRS Université de Toulouse (UPS), Toulouse, France.
authors.forename: Jun
authors.initials: J
authors.lastname: Liang
authors.affiliation: Laboratoire de Biologie Moléculaire Eucaryote (LBME), CNRS Université de Toulouse (UPS), Toulouse, France.
authors.forename: Gael
authors.initials: G
authors.lastname: Micas
authors.affiliation: Laboratoire de Biologie Moléculaire Eucaryote (LBME), CNRS Université de Toulouse (UPS), Toulouse, France.
authors.forename: Pauline
authors.initials: P
authors.lastname: Morillon
authors.affiliation: Mass-Spectrometry Facility, Institut de Génomique Fonctionnelle, Montpellier, France.
authors.forename: Serge
authors.initials: S
authors.lastname: Urbach
authors.affiliation: UMR444-Laboratoire de Génétique Cellulaire & GeT-PlaGe, INRA Genotoul, Auzeville, Toulouse, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Bouchez
authors.affiliation: MGX-Montpellier GenomiX, Institut de Génomique Fonctionnelle, Montpellier, France.
authors.forename: Dany
authors.initials: D
authors.lastname: Severac
authors.affiliation: Physics Department, Simon Fraser University (SFU), Burnaby, BC, Canada.
authors.forename: Eldon
authors.initials: E
authors.lastname: Emberly
authors.affiliation: Systems Biology Center, National Heart, Lung and Blood Institute National Institutes of Health (NIH), Bethesda, MD, USA.
authors.forename: Keji
authors.initials: K
authors.lastname: Zhao
authors.affiliation: Laboratoire de Biologie Moléculaire Eucaryote (LBME), CNRS Université de Toulouse (UPS), Toulouse, France cuvier@ibcg.biotoul.fr.
authors.forename: Olivier
authors.initials: O
authors.lastname: Cuvier
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002843:Chromatin / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism*; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism*; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000378:metabolism; D038101:Insulator Elements / Q000235:genetics*; D046228:Microarray Analysis; D008954:Models, Biological; D008969:Molecular Sequence Data; D009707:Nucleosomes / Q000502:physiology*; D025341:Principal Component Analysis; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017423:Sequence Analysis, RNA; D020798:Two-Hybrid System Techniques
chemical_list: C452919:BEAF-32 protein, Drosophila; D002843:Chromatin; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D005136:Eye Proteins; D006657:Histones; D009707:Nucleosomes; D011495:Histone-Lysine N-Methyltransferase; C527105:NSD protein, Drosophila
keywords: RNA splicing; chromatin barrier; higher‐order chromatin organization; nucleosome positioning; physical borders