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.

*DNA-Binding Proteins, *Homeodomain Proteins, Cations/pharmacology, Gene Rearrangement/drug effects/*genetics, Genes, Insect/genetics, Molecular Sequence Data, Proteins/*genetics, Sequence Analysis, DNA, animals, base sequence, insects, kinetics
hdl.handle.net/1765/9498
Cell
Erasmus MC: University Medical Center Rotterdam

van Gent, D.C. (1996). The RAG1 and RAG2 proteins establish the 12/23 rule in V(D)J recombination. Cell. Retrieved from http://hdl.handle.net/1765/9498