title: RUNX1-dependent RAG1 deposition instigates human TCR-δ locus rearrangement.
abstract: V(D)J recombination of TCR loci is regulated by chromatin accessibility to RAG1/2 proteins, rendering RAG1/2 targeting a potentially important regulator of lymphoid differentiation. We show that within the human TCR-α/δ locus, Dδ2-Dδ3 rearrangements occur at a very immature thymic, CD34(+)/CD1a(-)/CD7(+dim) stage, before Dδ2(Dδ3)-Jδ1 rearrangements. These strictly ordered rearrangements are regulated by mechanisms acting beyond chromatin accessibility. Importantly, direct Dδ2-Jδ1 rearrangements are prohibited by a B12/23 restriction and ordered human TCR-δ gene assembly requires RUNX1 protein, which binds to the Dδ2-23RSS, interacts with RAG1, and enhances RAG1 deposition at this site. This RUNX1-mediated V(D)J recombinase targeting imposes the use of two Dδ gene segments in human TCR-δ chains. Absence of this RUNX1 binding site in the homologous mouse Dδ1-23RSS provides a molecular explanation for the lack of ordered TCR-δ gene assembly in mice and may underlie differences in early lymphoid differentiation between these species.
authors.affiliation: Université Paris Descartes Sorbonne Cité, Institut Necker-Enfants Malades (INEM), Institut national de recherche médicale (INSERM) U1151, and Laboratory of Onco-Hematology, Assistance Publique-Hôpitaux de Paris (AP-HP), Hôpital Necker Enfants-Malades, 75015 Paris, France.
authors.forename: Agata
authors.initials: A
authors.lastname: Cieslak
authors.affiliation: Université Paris Descartes Sorbonne Cité, Institut Necker-Enfants Malades (INEM), Institut national de recherche médicale (INSERM) U1151, and Laboratory of Onco-Hematology, Assistance Publique-Hôpitaux de Paris (AP-HP), Hôpital Necker Enfants-Malades, 75015 Paris, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Le Noir
authors.affiliation: Université Paris Descartes Sorbonne Cité, Institut Necker-Enfants Malades (INEM), Institut national de recherche médicale (INSERM) U1151, and Laboratory of Onco-Hematology, Assistance Publique-Hôpitaux de Paris (AP-HP), Hôpital Necker Enfants-Malades, 75015 Paris, France.
authors.forename: Amélie
authors.initials: A
authors.lastname: Trinquand
authors.affiliation: Université Paris Descartes Sorbonne Cité, Institut Necker-Enfants Malades (INEM), Institut national de recherche médicale (INSERM) U1151, and Laboratory of Onco-Hematology, Assistance Publique-Hôpitaux de Paris (AP-HP), Hôpital Necker Enfants-Malades, 75015 Paris, France.
authors.forename: Ludovic
authors.initials: L
authors.lastname: Lhermitte
authors.affiliation: CNRS-Pierre Fabre USR3388, Epigenetic Targeting of Cancer (ETaC), and INSERM UMR1037, Cancer Research Center of Toulouse (CRCT), 31035 Toulouse, France.
authors.forename: Don-Marc
authors.initials: DM
authors.lastname: Franchini
authors.affiliation: Université Paris Descartes Sorbonne Cité, Institut Necker-Enfants Malades (INEM), Institut national de recherche médicale (INSERM) U1151, and Laboratory of Onco-Hematology, Assistance Publique-Hôpitaux de Paris (AP-HP), Hôpital Necker Enfants-Malades, 75015 Paris, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Villarese
authors.affiliation: Centre d'Immunologie de Marseille-Luminy (CIML), Aix-Marseille Université UM 2, INSERM UMR 1104, CNRS UMR 7280, 13288 Marseille, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Gon
authors.affiliation: Université Paris Descartes Sorbonne Cité, Institut Necker-Enfants Malades (INEM), Institut national de recherche médicale (INSERM) U1151, and Laboratory of Onco-Hematology, Assistance Publique-Hôpitaux de Paris (AP-HP), Hôpital Necker Enfants-Malades, 75015 Paris, France.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Bond
authors.affiliation: Université Paris Descartes Sorbonne Cité, Institut Necker-Enfants Malades (INEM), Institut national de recherche médicale (INSERM) U1151, and Laboratory of Onco-Hematology, Assistance Publique-Hôpitaux de Paris (AP-HP), Hôpital Necker Enfants-Malades, 75015 Paris, France.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Simonin
authors.affiliation: Technological Advances for Genomics and Clinics (TAGC), INSERM U1090, Université de la Méditerranée, 13288 Marseille, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Vanhille
authors.affiliation: Technological Advances for Genomics and Clinics (TAGC), INSERM U1090, Université de la Méditerranée, 13288 Marseille, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Vanhile
authors.affiliation: Université Paris-Descartes, Faculté de Médecine René Descartes, IFR94 and INSERM, U768, F-75015 Paris, France.
authors.forename: Christian
authors.initials: C
authors.lastname: Reimann
authors.affiliation: CIRI, International center for Infectiology Research, EVIR team, Université de Lyon, INSERM U1111, Lyon, France and Centre Méditerranéen de Médecine Moléculaire (C3M), team "contrôle métabolique des morts cellulaires" Inserm, U1065, 06204 Nice, France.
authors.forename: Els
authors.initials: E
authors.lastname: Verhoeyen
authors.affiliation: Assistance Publique-Hôpitaux de Paris (AP-HP), Hôpital Saint-Louis, Unité de Thérapie Cellulaire, Université Paris Diderot, Sorbonne Paris Cité, Inserm CICBT501 et UMR1160, Institut Universitaire d'Hématologie, 75010 Paris, France.
authors.forename: Jerome
authors.initials: J
authors.lastname: Larghero
authors.affiliation: Université Paris-Descartes, Faculté de Médecine René Descartes, IFR94 and INSERM, U768, F-75015 Paris, France.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Six
authors.affiliation: Technological Advances for Genomics and Clinics (TAGC), INSERM U1090, Université de la Méditerranée, 13288 Marseille, France.
authors.forename: Salvatore
authors.initials: S
authors.lastname: Spicuglia
authors.affiliation: Université Paris-Descartes, Faculté de Médecine René Descartes, IFR94 and INSERM, U768, F-75015 Paris, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: André-Schmutz
authors.affiliation: Department of Immunology, Erasmus MC, University Medical Center, 3016 Rotterdam, Netherlands.
authors.forename: Anton
authors.initials: A
authors.lastname: Langerak
authors.affiliation: Centre d'Immunologie de Marseille-Luminy (CIML), Aix-Marseille Université UM 2, INSERM UMR 1104, CNRS UMR 7280, 13288 Marseille, France.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Nadel
authors.affiliation: Université Paris Descartes Sorbonne Cité, Institut Necker-Enfants Malades (INEM), Institut national de recherche médicale (INSERM) U1151, and Laboratory of Onco-Hematology, Assistance Publique-Hôpitaux de Paris (AP-HP), Hôpital Necker Enfants-Malades, 75015 Paris, France.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Macintyre
authors.affiliation: Centre d'Immunologie de Marseille-Luminy (CIML), Aix-Marseille Université UM 2, INSERM UMR 1104, CNRS UMR 7280, 13288 Marseille, France vahid.asnafi@nck.aphp.fr payet@ciml.univ-mrs.fr.
authors.forename: Dominique
authors.initials: D
authors.lastname: Payet-Bornet
authors.affiliation: Université Paris Descartes Sorbonne Cité, Institut Necker-Enfants Malades (INEM), Institut national de recherche médicale (INSERM) U1151, and Laboratory of Onco-Hematology, Assistance Publique-Hôpitaux de Paris (AP-HP), Hôpital Necker Enfants-Malades, 75015 Paris, France vahid.asnafi@nck.aphp.fr payet@ciml.univ-mrs.fr.
authors.forename: Vahid
authors.initials: V
authors.lastname: Asnafi
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002454:Cell Differentiation; D002460:Cell Line; D050676:Core Binding Factor Alpha 2 Subunit / Q000378:metabolism*; D004247:DNA / Q000235:genetics / Q000378:metabolism; D016694:Gene Rearrangement, delta-Chain T-Cell Antigen Receptor; D057809:HEK293 Cells; D018398:Homeodomain Proteins / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D038041:Lymphopoiesis; D051379:Mice; D008969:Molecular Sequence Data; D013045:Species Specificity; D016176:T-Lymphocyte Subsets / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*; D045567:VDJ Recombinases / Q000378:metabolism
chemical_list: D050676:Core Binding Factor Alpha 2 Subunit; D018398:Homeodomain Proteins; C493728:RUNX1 protein, human; C064658:RAG-1 protein; D004247:DNA; D045567:VDJ Recombinases