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V(D)J recombination: in vitro coding joint formation.

F Weis-Garcia·E Besmer·D J Sawchuk·W Yu·Y Hu·S Cassard·M C Nussenzweig·P Cortes
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Abstract

Antigen receptor genes are assembled through a mechanism known as V(D)J recombination, which involves two different joining reactions: signal and coding joining. Formation of these joints is essential for antigen receptor assembly as well as maintaining chromosomal integrity. Here we report on a cell-free system for coding joint formation using deletion and inversion recombination substrates. In vitro coding joint formation requires RAG1, RAG2, and heat-labile factors present in the nuclear extract of nonlymphoid cells. Both inversion- and deletion-mediated coding joint reactions produce diverse coding joints, with deletions and P nucleotide addition. We also show that deletion-mediated coding joint formation follows the 12/23 rule and requires the catalytic subunit of DNA-dependent protein kinase.

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Cell NucleusCell-Free SystemDNADNA-Binding ProteinsHeLa CellsHomeodomain ProteinsHumansModels, GeneticNuclear ProteinsReceptors, AntigenRecombination, GeneticSubstrate SpecificityJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.

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F Weis-Garcia, E Besmer, D J Sawchuk, W Yu, Y Hu, S Cassard, M C Nussenzweig, P Cortes. 1997. V(D)J recombination: in vitro coding joint formation.. https://doi.org/10.1128/mcb.17.11.6379

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