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Intermolecular V(D)J recombination
1Howard Hughes Medical Institute, Section of Immunobiology, Yale University School of Medicine, New Haven, Connecticut 06520-8011, USA.
V(D)J recombination can occur between separate DNA molecules, forming coding joints. Linearization of substrates unexpectedly reduces coding joint formation, suggesting a "release and recapture" mechanism.
Area of Science:
- Immunology
- Molecular Biology
- Genetics
Background:
- V(D)J recombination is crucial for generating diverse antigen receptors in B and T lymphocytes.
- This process is implicated in chromosomal translocations, potentially via interchromosomal recombination.
Purpose of the Study:
- To investigate the V(D)J recombination machinery's capacity for intermolecular recombination.
- To explore the formation of coding joints in trans using plasmid substrates.
Main Methods:
- Investigated intermolecular V(D)J recombination between unlinked and catenated plasmids.
- Assessed the impact of substrate linearization on coding joint formation.
Main Results:
- V(D)J recombination occurred in trans between plasmid substrates, forming signal and coding joints.
- Substrate linearization significantly reduced coding joint formation without altering cis/trans status.
- No mechanistic block to trans coding joint formation was observed.
Conclusions:
- Proposed a "release and recapture" model for V(D)J recombination.
- Coding ends are frequently released from post-cleavage complexes and recaptured for joint formation.
- Chromosomal V(D)J recombination likely involves mechanisms preventing end release or facilitating recapture.
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