Intermolecular V(D)J recombination.
V(D)J recombination plays a prominent role in the generation of the antigen receptor repertoires of B and T lymphocytes. It is also likely to be involved in the formation of chromosomal translocations, some of which may result from interchromosomal recombination. We have investigated the potential of the V(D)J recombination machinery to perform intermolecular recombination between two plasmids, either unlinked or linked by catenation. In either case, recombination occurs in trans to yield signal and coding joints, and the results do not support the existence of a mechanistic block to the formation of coding joints in trans. Instead, we observe that linearization of the substrate, which does not alter the cis or trans status of the recombination signals, causes a specific and dramatic reduction in coding joint formation. This unexpected result leads us to propose a "release and recapture" model for V(D)J recombination in which coding ends are frequently released from the postcleavage complex and the efficiency of coding joint formation is influenced by the efficiency with which such ends are recaptured by the complex. This implies the existence of mechanisms, operative during recombination of chromosomal substrates, that act to prevent coding end release or to facilitate coding end recapture.
Tevelev
A
A
Howard Hughes Medical Institute, Section of Immunobiology, Yale University School of Medicine, New Haven, Connecticut 06520-8011, USA.
Schatz
D G
DG
D004264:DNA Topoisomerases, Type I / Q000378:metabolism; D004278:DNA, Superhelical / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D015321:Gene Rearrangement; D018398:Homeodomain Proteins / Q000378:metabolism; D008957:Models, Genetic; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000737:chemistry / Q000235:genetics*; D011946:Receptors, Antigen / Q000235:genetics*; D045522:Recombinases; D011995:Recombination, Genetic; D013379:Substrate Specificity; D019895:Transposases / Q000378:metabolism
D004278:DNA, Superhelical; D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D011946:Receptors, Antigen; D045522:Recombinases; C064655:V(D)J recombination activating protein 2; C064658:RAG-1 protein; D019895:Transposases; C084267:endodeoxyribonuclease XcmI; D015252:Deoxyribonucleases, Type II Site-Specific; D004264:DNA Topoisomerases, Type I