title: RAG1 and RAG2 form a stable postcleavage synaptic complex with DNA containing signal ends in V(D)J recombination.
abstract: During V(D)J recombination, RAG1 and RAG2 cleave DNA adjacent to highly conserved recombination signals, but nothing is known about the protein-DNA complexes that exist after cleavage. Using a properly regulated in vitro V(D)J cleavage system, together with nuclease sensitivity, mobility shift, and immunoprecipitation experiments, we provide evidence that a stable complex is formed postcleavage between synapsed recombination signals. This complex includes the proteins RAG1, RAG2, HMG-1 or the closely related HMG-2 protein, and the components of the DNA-dependent protein kinase. The existence of such a stable complex explains a number of in vivo observations and suggests that remodeling of postcleavage synaptic complexes is an important step in the resolution of signal ends in V(D)J recombination.
authors.affiliation: Department of Pharmacology, Yale University School of Medicine, New Haven, Connecticut 06520-8011, USA.
authors.forename: A
authors.initials: A
authors.lastname: Agrawal
authors.forename: D G
authors.initials: DG
authors.lastname: Schatz
mesh_terms: D000818:Animals; D004247:DNA / Q000032:analysis / Q000378:metabolism; D053938:DNA Fragmentation; D004254:DNA Nucleotidyltransferases / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000502:physiology; D004720:Endonucleases; D018398:Homeodomain Proteins; D016393:Lymphoma, B-Cell; D051379:Mice; D011233:Precipitin Tests; D011506:Proteins / Q000235:genetics*; D011817:Rabbits; D011995:Recombination, Genetic; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000201:enzymology; D045567:VDJ Recombinases
chemical_list: D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D011506:Proteins; C499309:RAG2 protein, Oryctolagus cuniculus; C499308:Rag2 protein, mouse; C064655:V(D)J recombination activating protein 2; C064658:RAG-1 protein; D004247:DNA; D004254:DNA Nucleotidyltransferases; D045567:VDJ Recombinases; D004720:Endonucleases