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J. Biol. Chem., Vol. 278, Issue 20, 18101-18109, May 16, 2003

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Assessing the Role of the T Cell Receptor  Gene Enhancer in Regulating Coding Joint Formation during V(D)J Recombination^*

Noëlle Mathieu, Salvatore Spicuglia, Sophie Gorbatch^§, Olivier Cabaud, Corinne Fernex, Christophe Verthuy, William M. Hempel^¶, Anne-Odile Hueber, and Pierre Ferrier^**

From the Centre d'Immunologie de Marseille-Luminy, INSERM, CNRS, Université de la Méditerranée, 13288 Marseille, France

To assess the role of the T cell receptor (TCR)  gene enhancer (E) in regulating the processing of VDJ recombinase-generated coding ends, we assayed TCR rearrangement of E-deleted (E) thymocytes in which cell death is inhibited via expression of a Bcl-2 transgene. Compared with E, E Bcl-2 thymocytes show a small accumulation of TCR standard recombination products, including coding ends, that involves the proximal D-J and V14 loci but not the distal 5' V genes. These effects are detectable in double negative pro-T cells, predominate in double positive pre-T cells, and correlate with regional changes in chromosomal structure during double negative-to-double positive differentiation. We propose that E, by driving long range nucleoprotein interactions and the control of locus expression and chromatin structure, indirectly contributes to the stabilization of coding ends within the recombination processing complexes. The results also illustrate E-dependent and -independent changes in chromosomal structure, suggesting distinct modes of regulation of TCR allelic exclusion depending on the position within the locus.

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