Initiation of V(D)J recombination in a cell-free system
- 16 June 1995
- journal article
- Published by Elsevier BV in Cell
- Vol. 81 (6), 925-934
- https://doi.org/10.1016/0092-8674(95)90012-8
Abstract
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: fulllength, 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
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