Identification of V(D)J recombination coding end intermediates in normal thymocytes.
Diversity of vertebrate antigen receptors is accomplished in large part by a somatic gene rearrangement process known as V(D)J recombination. The first step of the reaction appears to be the creation of a double strand break immediately between the recombination signal sequence (RSS) and the coding gene segment to generate a signal end and a coding end. Signal ends have been shown, both in vitro and in vivo, to be precise and blunt, while coding ends generated in vitro are covalently sealed hairpins. It has been difficult to document the existence of coding ends in vivo in normal lymphoid precursors, presumably because of their low abundance. To date, they have been identified in vivo only in a transformed pre-B cell line and in cells from the mutant scid mouse, where they largely conform to the hairpin structure found in vitro. Here, we identify T cell receptor J alpha gene coding ends in normal murine thymocytes. We demonstrate that these ends are processed, not blunt, and that most are not hairpin terminated, in sharp contrast to previous in vivo and in vitro observations. These results provide the first direct demonstration of this important intermediate of V(D)J recombination in normal lymphoid precursors and have implications for the mechanism of coding joint formation in vivo.
Livák
F
F
Section of Immunology, Yale Medical School, New Haven, CT 06520-8011, USA.
Schatz
D G
DG
D000818:Animals; D004260:DNA Repair; D015321:Gene Rearrangement; D007133:Immunoglobulin Joining Region / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D051379:Mice; D016513:Mice, SCID; D021382:Protein Sorting Signals / Q000235:genetics; D011995:Recombination, Genetic; D013950:Thymus Gland / Q000166:cytology / Q000378:metabolism*
D007133:Immunoglobulin Joining Region; D007135:Immunoglobulin Variable Region; D021382:Protein Sorting Signals