title: Replacement of Imu-Cmu intron by NeoR gene alters Imu germ-line expression but has no effect on V(D)J recombination.
abstract: The NeoR gene has often been used to unravel the mechanisms underlying long-range interactions between promoters and enhancers during V(D)J assembly and class switch recombination (CSR) in the immunoglobulin heavy chain (IgH) locus. This approach led to the notion that CSR is regulated through competition of germ-line (GL) promoters for activities displayed by the 3' regulatory region (3'RR). This polarized long-range effect of the 3'RR is disturbed upon insertion of NeoR gene in the IgH constant (C(H)) region, where only GL transcription derived from upstream GL promoters is impaired. In the context of V(D)J recombination, replacement of Emu enhancer or Emu core enhancer (cEmu) by NeoR gene fully blocked V(D)J recombination and mu0 GL transcription which originates 5' of DQ52 and severely diminished Imu GL transcription derived from Emu/Imu promoter, suggesting a critical role for cEmu in the regulation of V(D)J recombination and of mu0 and Imu expression. Here we focus on the effect of NeoR gene on mu0 and Imu GL transcription in a mouse line in which the Imu-Cmu intron was replaced by a NeoR gene in the sense-orientation. B cell development was characterized by a marked but incomplete block at the pro-B cell stage. However, V(D)J recombination was unaffected in sorted pro-B and pre-B cells excluding an interference with the accessibility control function of Emu. mu0 GL transcription initiation was relatively normal but the maturation step seemed to be affected most likely through premature termination at NeoR polyadenylation sites. In contrast, Imu transcription initiation was impaired suggesting an interference of NeoR gene with the IgH enhancers that control Imu expression. Surprisingly, in stark contrast with the NeoR effect in the C(H) region, LPS-induced NeoR expression restored Imu transcript levels to normal. The data suggest that Emu enhancer may be the master control element that counteracts the down-regulatory "Neo effect" on Imu expression upon LPS stimulation. More importantly, they reveal a complex and developmentally regulated interplay between IgH enhancers in the control of Imu expression.
authors.affiliation: CNRS UMR 5089, Institut de Pharmacologie et de Biologie Structurale, Equipe Instabilité génétique et régulation transcriptionnelle, 205 route de Narbonne, 31077 Toulouse, France.
authors.forename: Dania
authors.initials: D
authors.lastname: Haddad
authors.forename: Hei-Lanne
authors.initials: HL
authors.lastname: Dougier
authors.forename: Nathalie
authors.initials: N
authors.lastname: Laviolette
authors.forename: Nadine
authors.initials: N
authors.lastname: Puget
authors.forename: Ahmed Amine
authors.initials: AA
authors.lastname: Khamlichi
mesh_terms: D000818:Animals; D004742:Enhancer Elements, Genetic / Q000235:genetics / Q000276:immunology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics / Q000276:immunology; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain / Q000235:genetics* / Q000276:immunology; D005798:Genes, Bacterial / Q000235:genetics* / Q000276:immunology; D007127:Immunoglobulin Constant Regions / Q000235:genetics / Q000276:immunology; D007148:Immunoglobulin mu-Chains / Q000235:genetics* / Q000276:immunology; D007438:Introns / Q000235:genetics* / Q000276:immunology; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D008817:Mice, Mutant Strains; D016254:Mutagenesis, Insertional; D054448:Precursor Cells, B-Lymphoid / Q000276:immunology; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000276:immunology; D040641:Quantitative Trait Loci / Q000235:genetics* / Q000276:immunology; D027041:Somatic Hypermutation, Immunoglobulin / Q000235:genetics* / Q000276:immunology; D014158:Transcription, Genetic / Q000187:drug effects / Q000235:genetics / Q000276:immunology
chemical_list: D007127:Immunoglobulin Constant Regions; D007148:Immunoglobulin mu-Chains; D008070:Lipopolysaccharides