Abstract
Allelic exclusion of Vβ-to-DJβ recombination depends on asynchronous rearrangement of alleles of the gene encoding T cell receptor β in double-negative thymocytes and feedback inhibition that is maintained in double-positive thymocytes. Feedback is thought to be enforced through downregulation of Vβ accessibility. In an attempt to override this negative regulation, we introduced the enhancer of the gene encoding T cell receptor α into the Vβ gene cluster downstream of Vβ12. In double-negative thymocytes, the introduced enhancer had no measurable effect on accessibility, but Vβ12 rearrangement was stimulated and Vβ12 allelic exclusion was partially subverted. In contrast, double-positive thymocytes showed increased Vβ transcription and accessibility, but feedback inhibition of Vβ-to-DJβ recombination remained intact. Our results indicate additional regulatory constraints on Vβ-to-DJβ recombination that operate beyond the accessibility barrier.
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References
Jung, D. & Alt, F.W. Unraveling V(D)J recombination: Insights into gene regulation. Cell 116, 299–311 (2004).
Bassing, C.H., Swat, W. & Alt, F.W. The mechanism and regulation of chromosomal V(D)J recombination. Cell 109, S45–S55 (2002).
Krangel, M.S. Gene segment selection in V(D)J recombination: accessibility and beyond. Nat. Immunol. 4, 624–630 (2003).
Khor, B. & Sleckman, B.P. Allelic exclusion at the TCRβ locus. Curr. Opin. Immunol. 14, 230–234 (2002).
Mostoslavsky, R., Alt, F.W. & Rajewsky, K. The lingering enigma of the allelic exclusion mechanism. Cell 118, 539–544 (2004).
Bergman, Y., Fisher, A. & Cedar, H. Epigenetic mechanisms that regulate antigen receptor gene expression. Curr. Opin. Immunol. 15, 176–181 (2003).
Schlissel, M.S. Regulating antigen-receptor gene assembly. Nat. Rev. Immunol. 3, 890–899 (2003).
Narlikar, G.J., Fan, H.Y. & Kingston, R.E. Cooperation between complexes that regulate chromatin structure and transcription. Cell 108, 475–487 (2002).
Golding, A., Chandler, S., Ballestar, E., Wolffe, A.P. & Schlissel, M.S. Nucleosome structure completely inhibits in vitro cleavage by the V(D)J recombinase. EMBO J. 18, 3712–3723 (1999).
Kwon, J., Imbalzano, A.N., Matthews, A. & Oettinger, M.A. Accessiblity of nucleosomal DNA to V(D)J cleavage is modulated by RSS positioning and HMG1. Mol. Cell 2, 829–839 (1998).
Stanhope-Baker, P., Hudson, K.M., Shaffer, A.L., Constantinescu, A. & Schlissel, M.S. Cell type-specific chromatin structure determines the targeting of V(D)J recombinase activity in vitro. Cell 85, 887–897 (1996).
Baumann, M., Mamais, A., McBlane, F., Xiao, H. & Boyes, J. Regulation of V(D)J recombination by nucleosome positioning at recombination signal sequences. EMBO J. 22, 5197–5207 (2003).
McMurry, M.T. & Krangel, M.S. A role for histone acetylation in the developmental regulation of V(D)J recombination. Science 287, 495–498 (2000).
Mathieu, N., Hempel, W.M., Spicuglia, S., Verthuy, C. & Ferrier, P. Chromatin remodeling by the T cell receptor (TCR)-β gene enhancer during early T cell development: implications for the control of TCR-β locus recombination. J. Exp. Med. 192, 625–636 (2000).
Goldmit, M., Schlissel, M., Cedar, H. & Bergman, Y. Differential accessibility at the κ chain locus plays a role in allelic exclusion. EMBO J. 21, 5255–5261 (2002).
Ji, Y.H., Zhang, J.M., Lee, A.I., Cedar, H. & Bergman, Y. A multistep mechanism for the activation of rearrangement in the immune system. Proc. Natl. Acad. Sci. USA 100, 7557–7562 (2003).
Morshead, K.B., Ciccone, D.N., Taverna, S.D., Allis, C.D. & Oettinger, M.A. Antigen receptor loci poised for V(D)J rearrangement are broadly associated with BRG1 and flanked by peaks of histone H3 dimethylated at lysine 4. Proc. Natl. Acad. Sci. USA 100, 11577–11582 (2003).
Osipovich, O. et al. Targeted inhibition of V(D)J recombination by a histone methyltransferase. Nat. Immunol. 5, 309–316 (2004).
Mostoslavsky, R. et al. Asynchronous replication and allelic exclusion in the immune system. Nature 414, 221–225 (2001).
Liang, H.-E. et al. The “dispensable” portion of RAG2 is necessary for efficient V-to-DJ rearrangement during B and T cell development. Immunity 17, 639–651 (2002).
Liang, H.E., Cado, D. & Schlissel, M. Variegated transcriptional activation of the immunoglobulin kappa locus in pre-B cells contribute to the allelic exclusion of light-chain expression. Cell 118, 19–29 (2004).
Michie, A. & Zuniga-Pflucker, J. Regulation of thymocyte differentiation: pre-TCR signals and β-selection. Semin. Immunol. 14, 311–323 (2002).
Senoo, M. & Shinkai, Y. Regulation of Vβ germline transcription in RAG-deficient mice by the CD3ε-mediated signals: implications of Vβ transcriptional regulation in TCR β allelic exclusion. Int. Immunol. 10, 553–560 (1998).
Tripathi, R., Jackson, A. & Krangel, M.S. A change in the structure of Vβ chromatin associated with TCRβ allelic exclusion. J. Immunol. 168, 2316–2324 (2002).
Mathieu, N. et al. Assessing the role of the T cell receptor β gene enhancer in regulating coding joint formation during V(D)J recombination. J. Biol. Chem. 278, 18101–18109 (2003).
Chattopadhyay, S., Whitehurst, C.E., Schwenk, F. & Chen, J. Biochemical and functional analysis of chromatin changes at the TCR-β gene locus during CD4−CD8− to CD4+CD8+ thymocyte differentiation. J. Immunol. 160, 1256–1267 (1998).
Whitehurst, C.E., Chattopadhyay, S. & Chen, J. Control of V(D)J recombinational accessibility of the Dβ1 gene segment at the TCRβ locus by a germline promoter. Immunity 10, 313–322 (1999).
Maes, J. et al. Chromatin remodeling at the Ig loci prior to V(D)J recombination. J. Immunol. 167, 866–874 (2001).
Johnson, K., Angelin-Duclos, C., Park, S. & Calame, K.L. Changes in histone acetylation are associated with differences in accessibility of VH gene segments to V-DJ recombination during B-cell ontogeny and development. Mol. Cell. Biol. 23, 2438–2450 (2003).
Chowdhury, D. & Sen, R. Transient IL-7/IL-7R signaling provides a mechanism for feedback inhibition of immunoglobulin heavy chain gene rearrangements. Immunity 18, 229–241 (2003).
Ryu, C.J. et al. The T-cell receptor β variable gene promoter is required for efficient Vβ rearrangement but not allelic exclusion. Mol. Cell. Biol. 24, 7015–7023 (2004).
Hernandez-Munain, C., Sleckman, B.P. & Krangel, M.S. A developmental switch from TCRδ enhancer to TCRα enhancer function during thymocyte maturation. Immunity 10, 723–733 (1999).
Capone, M. et al. TCRβ and TCRα gene enhancers confer tissue- and stage-specificity on V(D)J recombination events. EMBO J. 12, 4335–4346 (1993).
Senoo, M. et al. Limited effect of chromatin remodeling on Dβ-to-Jβ recombination in CD4+CD8+ thymocyte: implications for a new aspect in the regulation of TCRβ gene recombination. Int. Immunol. 13, 1405–1414 (2001).
Bassing, C.H. et al. Recombination signal sequences restrict chromosomal V(D)J recombination beyond the 12/23 rule. Nature 405, 583–586 (2000).
Davodeau, F. et al. Dual T cell receptor β chain expression on human T lymphocytes. J. Exp. Med. 181, 1391–1398 (1995).
Padovan, E. et al. Normal T lymphocytes can express two different T cell receptor β chains: implications for the mechanism of allelic exclusion. J. Exp. Med. 181, 1587–1591 (1995).
King, A.G., Kondo, M., Scherer, D.C. & Weissman, I.L. Lineage infidelity in myeloid cells with TCR gene rearrangement: A latent developmental potential of proT cells revealed by ectopic cytokine receptor signaling. Proc. Natl. Acad. Sci. USA 99, 4508–4513 (2002).
Spicuglia, S. et al. TCRα enhancer activation occurs via a conformational change of a pre-assembled nucleoprotein complex. EMBO J. 19, 2034–2045 (2000).
Ryu, C.J. et al. The T cell receptor β enhancer promotes access and pairing of Dβ and Jβ gene segments during V(D)J recombination. Proc. Natl. Acad. Sci. USA 100, 13465–13470 (2003).
Senoo, M. et al. Increase of TCR Vβ accessibility within Eβ regulatory region influences its recombination frequency but not allelic exclusion. J. Immunol. 171, 829–835 (2003).
Hesslein, D.G. et al. Pax5 is required for recombination of transcribed, acetylated, 5′ IgH V gene segments. Genes Dev. 17, 37–42 (2003).
Fuxa, M. et al. Pax5 induces V-to-DJ rearrangements and locus contraction of the immunoglobulin heavy-chain gene. Genes Dev. 18, 411–422 (2004).
Roldán, E. et al. Locus 'decontraction' and centromeric recruitment contribute to allelic exclusion of the immunoglobulin heavy-chain gene. Nat. Immunol. 6, 31–41 (2005).
Shinkai, Y. et al. Restoration of T cell development in RAG-2-deficient mice by functional TCR transgenes. Science 259, 822–825 (1993).
Roberts, J.L., Lauzurica, P. & Krangel, M.S. Developmental regulation of VDJ recombination by the core fragment of the T cell receptor α enhancer. J. Exp. Med. 185, 131–140 (1997).
Boyd, K.E., Wells, J., Gutman, J., Bartley, S.M. & Farnham, P.J. c-Myc target gene specificity is determined by a post-DNA-binding mechanism. Proc. Natl. Acad. Sci. USA 95, 13887–13892 (1998).
Boyes, J. & Felsenfeld, G. Tissue-specific factors additively increase the probability of the all-or-none formation of a hypersensitive site. EMBO J. 15, 2496–2507 (1996).
McMurry, M.T., Hernandez-Munain, C., Lauzurica, P. & Krangel, M.S. Enhancer control of local accessibility to V(D)J recombinase. Mol. Cell. Biol. 17, 4553–4561 (1997).
Acknowledgements
We thank L. Martinek of the Duke University Cancer Center Flow Cytometry Facility for help with cell sorting and analysis; T. Wehrly and H. Boutrid for technical assistance; and Y. Zhuang, Y.-W. He and G. Kelsoe for critical review of the manuscript. Supported by the National Institutes of Health (AI49934 to M.S.K. and NIAID-T32 AI052077 to H.D.K.) and National Science Foundation (A.J.).
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Supplementary information
Supplementary Fig. 1
DNase Ihypersensitivity of Eαi in DN and DP thymocytes of EαKI mice. (PDF 119 kb)
Supplementary Fig. 2
Strategy for restriction endonuclease accessibility assay. (PDF 45 kb)
Supplementary Table 1
Oligonucleotides used as PCR primers or radiolabeled probes. (PDF 44 kb)
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Jackson, A., Kondilis, H., Khor, B. et al. Regulation of T cell receptor β allelic exclusion at a level beyond accessibility. Nat Immunol 6, 189–197 (2005). https://doi.org/10.1038/ni1157
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DOI: https://doi.org/10.1038/ni1157
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