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1091-649010622009Jan13Proceedings of the National Academy of Sciences of the United States of AmericaProc Natl Acad Sci U S ABiallelic, ubiquitous transcription from the distal germline Ig{kappa} locus promoter during B cell development.522527522-710.1073/pnas.0808895106Allelic exclusion of Ig gene expression is necessary to limit the number of functional receptors to one per B cell. The mechanism underlying allelic exclusion is unknown. Because germline transcription of Ig and TCR loci is tightly correlated with rearrangement, we created two novel knock-in mice that report transcriptional activity of the Jkappa germline promoters in the Igkappa locus. Analysis of these mice revealed that germline transcription is biallelic and occurs in all pre-B cells. Moreover, we found that the two germline promoters in this region are not equivalent but that the distal promoter accounts for the vast majority of observed germline transcript in pre-B cells while the activity of the proximal promoter increases later in development. Allelic exclusion of the Igkappa locus thus occurs at the level of rearrangement, but not germline transcription.AminRupesh HRHDepartment of Molecular and Cell Biology, 439 Life Science Addition, University of California, Berkeley, CA 94720, USA.CadoDraganaDNollaHectorHHuangDanDShintonSusan ASAZhouYanYHardyRichard RRRSchlisselMark SMSengR01 HL048702HLNHLBI NIH HHSUnited StatesR01 HL48702HLNHLBI NIH HHSUnited StatesJournal ArticleResearch Support, N.I.H., Extramural20081230
United StatesProc Natl Acad Sci U S A75058760027-84240Immunoglobulin kappa-ChainsIMAllelesAnimalsB-LymphocytescytologyimmunologyGene Knock-In TechniquesGene RearrangementHumansImmunoglobulin kappa-ChainsgeneticsMiceMice, TransgenicPrecursor Cells, B-LymphoidcytologyimmunologyPromoter Regions, GeneticTranscription, GeneticThe authors declare no conflict of interest.20091190200911902009214902009713ppublish19116268PMC262673610.1073/pnas.08088951060808895106Schlissel MS. Regulating antigen-receptor gene assembly. Nat Rev Immunol. 2003;3:890–899.14668805Burnet FM, Fenner F. The Production of Antibodies. 2nd Ed. Melbourne: Macmillan; 1949. p. viii.Corcoran AE. Immunoglobulin locus silencing and allelic exclusion. Semin Immunol. 2005;17:141–154.15737575Mostoslavsky R, et al. Asynchronous replication and allelic exclusion in the immune system. Nature. 2001;414:221–225.11700561Liang HE, Hsu LY, Cado D, Schlissel MS. Variegated transcriptional activation of the immunoglobulin kappa locus in pre-b cells contributes to the allelic exclusion of light-chain expression. Cell. 2004;118:19–29.15242641Mostoslavsky R, Alt FW, Rajewsky K. The lingering enigma of the allelic exclusion mechanism. Cell. 2004;118:539–544.15339659Cobb RM, Oestreich KJ, Osipovich OA, Oltz EM. Accessibility control of V(D)J recombination. Adv Immunol. 2006;91:45–109.16938538Schlissel MS, Baltimore D. Activation of immunoglobulin kappa gene rearrangement correlates with induction of germline kappa gene transcription. Cell. 1989;58:1001–1007.2505932Abarrategui I, Krangel MS. Regulation of T cell receptor-alpha gene recombination by transcription. Nat Immunol. 2006;7:1109–1115.16936730Leclercq L, Butkeraitis P, Reth M. A novel germ-line JK transcript starting immediately upstream of JK1. Nucleic Acids Res. 1989;17:6809–6819.PMC3184142506531Martin DJ, van Ness BG. Initiation and processing of two kappa immunoglobulin germ line transcripts in mouse B cells. Mol Cell Biol. 1990;10:1950–1958.PMC3605412109186Van Ness BG, et al. Transcription of the unrearranged mouse C kappa locus: Sequence of the initiation region and comparison of activity with a rearranged V kappa-C kappa gene. Cell. 1981;27:593–602.6101210Cocea L, et al. A targeted deletion of a region upstream from the Jkappa cluster impairs kappa chain rearrangement in cis in mice and in the 103/bcl2 cell line. J Exp Med. 1999;189:1443–1450.PMC219306110224284Ferradini L, et al. Rearrangement-enhancing element upstream of the mouse immunoglobulin kappa chain J cluster. Science. 1996;271:1416–1420.8596914Singh N, Bergman Y, Cedar H, Chess A. Biallelic germline transcription at the kappa immunoglobulin locus. J Exp Med. 2003;197:743–750.PMC219385612629064Moebius U, Pallai P, Harrison SC, Reinherz EL. Delineation of an extended surface contact area on human CD4 involved in class II major histocompatibility complex binding. Proc Natl Acad Sci USA. 1993;90:8259–8263.PMC473288367491Casellas R, et al. Contribution of receptor editing to the antibody repertoire. Science. 2001;291:1541–1544.11222858Pikaart MJ, Recillas-Targa F, Felsenfeld G. Loss of transcriptional activity of a transgene is accompanied by DNA methylation and histone deacetylation and is prevented by insulators. Genes Dev. 1998;12:2852–2862.PMC3171659744862Schaefer BC. Revolutions in rapid amplification of cDNA ends: New strategies for polymerase chain reaction cloning of full-length cDNA ends. Anal Biochem. 1995;227:255–273.7573945Schlissel MS. Regulation of activation and recombination of the murine Igkappa locus. Immunol Rev. 2004;200:215–223.15242407Stanhope-Baker P, Hudson KM, Shaffer AL, Constantinescu A, Schlissel MS. Cell type-specific chromatin structure determines the targeting of V(D)J recombinase activity in vitro. Cell. 1996;85:887–897.8681383Goldmit M, Schlissel M, Cedar H, Bergman Y. Differential accessibility at the kappa chain locus plays a role in allelic exclusion. EMBO J. 2002;21:5255–5261.PMC12904012356741Goldmit M, et al. Epigenetic ontogeny of the Igk locus during B cell development. Nat Immunol. 2005;6:198–203.15619624Mostoslavsky R, et al. Kappa chain monoallelic demethylation and the establishment of allelic exclusion. Genes Dev. 1998;12:1801–1811.PMC3169089637682Kosak ST, et al. Subnuclear compartmentalization of immunoglobulin loci during lymphocyte development. Science. 2002;296:158–162.11935030Gilbert N, et al. Chromatin architecture of the human genome: Gene-rich domains are enriched in open chromatin fibers. Cell. 2004;118:555–566.15339661Schneider R, Grosschedl R. Dynamics and interplay of nuclear architecture, genome organization, and gene expression. Genes Dev. 2007;21:3027–3043.18056419Rumfelt LL, Zhou Y, Roweley BM, Shinton SA, Hardy RR. Lineage specification and plasticity in CD19- early B cell precursors. J Exp Med. 2006;203:675–687.PMC211824116505143Hardy RR, Shinton SA. Characterization of B lymphopoiesis in mouse bone marrow and spleen. Methods Mol Biol. 2004;271:1–24.15146109