title: Regions of RAG1 protein critical for V(D)J recombination.
abstract: The products of the recombination activating genes RAG1 and RAG2 are essential for activating V(D)J recombination, and thus are indispensable for the production of functional and diverse antigen receptors. To investigate the function of RAG1, we have tested a series of insertion and substitution mutation for their ability to induce V(D)J rearrangement on both deletional and inversional plasmid substrates. With these substrates we were also able to assess the effects of these mutations on both coding and signal joint formation, and to show that any one mutant affected all these reactions similarly. As defined previously, the core active regions of RAG1 and RAG2 permit the deletion of 40% and 25%, respectively, of well-conversed sequence. We show here that this "dispensable" region of RAG1 is not necessary for coding joint formation or recombination of an integrated substrate, and this portion is not functionally redundant with the "dispensable" region of RAG2. Recombination with these core regions is also still subject to the 12/23 joining rule. Further, the minimal essential core region of RAG1 can be located within an even smaller portion of the gene.
authors.affiliation: Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, USA.
authors.forename: S A
authors.initials: SA
authors.lastname: Kirch
authors.forename: P
authors.initials: P
authors.lastname: Sudarsanam
authors.forename: M A
authors.initials: MA
authors.lastname: Oettinger
mesh_terms: D001483:Base Sequence; D004254:DNA Nucleotidyltransferases / Q000378:metabolism; D004268:DNA-Binding Proteins; D004926:Escherichia coli; D015322:Gene Rearrangement, B-Lymphocyte; D006367:HeLa Cells; D018398:Homeodomain Proteins; D006801:Humans; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016297:Mutagenesis, Site-Directed; D009687:Nuclear Proteins; D010957:Plasmids / Q000235:genetics; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017384:Sequence Deletion; D013329:Structure-Activity Relationship; D014162:Transfection; D045567:VDJ Recombinases
chemical_list: D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D009687:Nuclear Proteins; D011506:Proteins; C499311:RAG2 protein, human; D011993:Recombinant Fusion Proteins; C064655:V(D)J recombination activating protein 2; C064658:RAG-1 protein; D004254:DNA Nucleotidyltransferases; D045567:VDJ Recombinases