In vitro V(D)J recombination: signal joint formation.
basic_science · Level V
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- Record sourced from PubMed, PMID 8943051.
- Also identified by PMC identifier 19485.
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Abstract
The first step of V(D)J recombination, specific cleavage at the recombination signal sequence (RSS), can be carried out by the recombination activating proteins RAG1 and RAG2. In vivo, the cleaved coding and signal ends must be rejoined to generate functional antigen receptors and maintain chromosomal integrity. We have investigated signal joint formation using deletion and inversion substrates in a cell free system. RAG1 and RAG2 alone or in combination were unable to generate signal joints. However, RAG1 and RAG2 complemented with nuclear extracts were able to recombine an extrachromosomal substrate and form precise signal joints. The in vitro reaction resembled authentic V(D)J recombination in being Ku-antigen-dependent.
Medical subject headings
- Antigens, Nuclear
- DNA Helicases
- DNA Nucleotidyltransferases
- Homeodomain Proteins
- Immunoglobulin Joining Region
- Immunoglobulin Variable Region
- Proteins