Initiation of V(D)J recombination in a cell-free system

Dik C. van Gent, J. Fraser McBlane, Dale A. Ramsden, Moshe J. Sadofsky, Joanne E. Hesse, Martin Gellert

Research output: Contribution to journalArticlepeer-review

264 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)925-934
Number of pages10
JournalCell
Volume81
Issue number6
DOIs
StatePublished - Jun 16 1995
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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