AI Article Synopsis
- Pre-B cell nuclear extracts can perform specific DNA cuts at recombination signal sequences, producing consistent cleavage products found in thymocytes.
- The presence of a complete signal sequence is necessary for this cleavage to occur, and recombinant RAG1 protein enhances this activity in extracts lacking RAG1.
- Cleavage activity in fractionated extracts correlates with the presence of RAG2, indicating that both RAG1 and RAG2 are essential components of the V(D)J recombinase complex.
Article 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: 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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| http://dx.doi.org/10.1016/0092-8674(95)90012-8 | DOI Listing |
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