Initiation of V (D) J recombination in a cell-free system
DC van Gent, JF McBlane, DA Ramsden, MJ Sadofsky… - Cell, 1995 - cell.com
DC van Gent, JF McBlane, DA Ramsden, MJ Sadofsky, JE Hesse, M Gellert
Cell, 1995•cell.comSummary 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 …
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 …
Summary
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.
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