The RAG1 and RAG2 proteins establish the 12/23 rule in V(D)J recombination

Cell. 1996 Apr 5;85(1):107-13. doi: 10.1016/s0092-8674(00)81086-7.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cations / pharmacology
  • DNA-Binding Proteins*
  • Gene Rearrangement / drug effects
  • Gene Rearrangement / genetics*
  • Genes, Insect / genetics
  • Homeodomain Proteins*
  • Insecta
  • Kinetics
  • Molecular Sequence Data
  • Proteins / genetics*
  • Sequence Analysis, DNA


  • Cations
  • DNA-Binding Proteins
  • Homeodomain Proteins
  • Proteins
  • V(D)J recombination activating protein 2
  • RAG-1 protein