title: Duality of enhancer functioning mode revealed in a reduced TCR beta gene enhancer knockin mouse model.
abstract: The TCRbeta gene enhancer (Ebeta) commands TCRbeta gene expression through the lifespan of T lymphocytes. Genetic and molecular studies have implied that in early thymocytes, Ebeta directs chromatin opening over the Dbeta-Jbeta-Cbeta domains and triggers initial Dbeta-Jbeta recombination. In mature T cells, Ebeta is required for expression of the assembled TCRbeta gene. Whether these separate activities rely on distinct Ebeta regulatory sequences and involve differing modes of activation is unclear. Using gene targeting in mouse embryonic stem cells, we replaced Ebeta by a conserved core fragment (Ebeta169). We found that Ebeta169-carrying alleles were capable of sustaining beta gene expression and the development of mature T cells in homozygous knockin mice. Surprisingly, these procedures and underlying molecular transactions were affected to a wide range of degrees depending on the developmental stage. Early thymocytes barely achieved Dbeta-Jbeta germline transcription and recombination. In contrast, T cells displayed substantial though heterogeneous levels of VDJ-rearranged TCRbeta gene expression. Our results have implications regarding enhancer function in cells of the adaptive immune system and, potentially, TCRbeta gene recombination and allelic exclusion.
authors.affiliation: Centre d'Immunologie de Marseille-Luminy, Université Aix Marseille, Marseille, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Bonnet
authors.forename: Fang
authors.initials: F
authors.lastname: Huang
authors.forename: Touati
authors.initials: T
authors.lastname: Benoukraf
authors.forename: Olivier
authors.initials: O
authors.lastname: Cabaud
authors.forename: Christophe
authors.initials: C
authors.lastname: Verthuy
authors.forename: Anaelle
authors.initials: A
authors.lastname: Boucher
authors.forename: Sébastien
authors.initials: S
authors.lastname: Jaeger
authors.forename: Pierre
authors.initials: P
authors.lastname: Ferrier
authors.forename: Salvatore
authors.initials: S
authors.lastname: Spicuglia
mesh_terms: D000483:Alleles; D000818:Animals; D001665:Binding Sites / Q000235:genetics / Q000276:immunology; D004742:Enhancer Elements, Genetic / Q000235:genetics / Q000276:immunology*; D055879:Gene Knock-In Techniques; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor / Q000276:immunology; D006825:Hybridomas; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008817:Mice, Mutant Strains; D008822:Mice, Transgenic; D018448:Models, Immunological; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics*; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*
chemical_list: D007135:Immunoglobulin Variable Region; D016693:Receptors, Antigen, T-Cell, alpha-beta