Initiation of V(D)J recombination in a cell-free system
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.
|Keywords||*DNA-Binding Proteins, *Genes, Immunoglobulin, *Homeodomain Proteins, *Recombination, Genetic, B-Lymphocytes/immunology, DNA Primers/genetics, DNA/genetics, Molecular Sequence Data, Receptors, Antigen, T-Cell/*genetics, animals, base sequence, cell line, cell-free system|
van Gent, D.C., McBlane, J.F., Ramsden, D.A., Sadokfsky, M.J., Hesse, J.E., & Gellert, M.. (1995). Initiation of V(D)J recombination in a cell-free system. Cell. Retrieved from http://hdl.handle.net/1765/9489