Cells performing V(D)J recombination make specific cuts in DNA at recombination signal sequences. Here, we show that nuclear extracts of pre-B cell lines carry out this specific cleavage. The products of cleavage are the same as found previously in thymocytes: full-length, blunt, 5'-phosphorylated signal ends, and covalently sealed (hairpin) coding ends. A complete signal sequence is required. Recombinant RAG1 protein greatly increases activity and complements an inactive extract from a RAG1 (-/-) pre-B cell line. When the extracts are fractionated, cleavage activity correlates with the presence of RAG2 protein. These results suggest that RAG1 and RAG2 are components of the V(D)J recombinase.

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Erasmus MC: University Medical Center Rotterdam

van Gent, D., McBlane, J. F., Ramsden, D. A., Sadokfsky, M. J., Hesse, J., & Gellert, M. (1995). Initiation of V(D)J recombination in a cell-free system. Cell. Retrieved from http://hdl.handle.net/1765/9489