The RAG1 and RAG2 proteins establish the 12/23 rule in V(D)J recombination.
V(D)J recombination requires a pair of signal sequences with spacer lengths of 12 and 23 base pairs. Cleavage by the RAG1 AND RAG2 proteins was previously shown to demand only a single signal sequence. Here, we established conditions where 12- and 23-spacer signal sequences are both necessary for cleavage. Coupled cutting at both sites requires only the RAG1 and RAG2 proteins, but depends on the metal ion. In Mn2+, a single signal sequence supports efficient double strand cleavage, but cutting in Mg2+ requires two signal sequences and is best with the canonical 12/23 pair. Thus, the RAG proteins determine both aspects of the specificity of V(D)J recombination, the recognition of a single signal sequence and the correct 12/23 coupling in a pair of signals.
van Gent
D C
DC
Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.
Ramsden
D A
DA
Gellert
M
M
D000818:Animals; D001483:Base Sequence; D002412:Cations / Q000494:pharmacology; D004268:DNA-Binding Proteins; D015321:Gene Rearrangement / Q000187:drug effects / Q000235:genetics*; D017344:Genes, Insect / Q000235:genetics; D018398:Homeodomain Proteins; D007313:Insecta; D007700:Kinetics; D008969:Molecular Sequence Data; D011506:Proteins / Q000235:genetics*; D017422:Sequence Analysis, DNA
D002412:Cations; D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D011506:Proteins; C064655:V(D)J recombination activating protein 2; C064658:RAG-1 protein