RAG1 Mediates Signal Sequence Recognition and Recruitment of RAG2 in V(D)J Recombination

Recent studies have demonstrated that DNA cleavage during V(D)J recombination is mediated by the RAG1 and RAG2 proteins. These proteins must therefore bind to the recombination signals, but the specific binding interaction has been difficult to study in vitro. Here, we use an in vivo one-hybrid DNA...

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Bibliographic Details
Published in:Cell 1996-10, Vol.87 (2), p.253-262
Main Authors: Difilippantonio, Michael J, McMahan, Catherine J, Eastman, Quinn M, Spanopoulou, Eugenia, Schatz, David G
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Cell Line
DNA Nucleotidyltransferases - genetics
DNA-Binding Proteins - physiology
Genes, Immunoglobulin
Homeodomain Proteins
Humans
Macromolecular Substances
Molecular Sequence Data
Nuclear Proteins
Protein Binding
Proteins - physiology
Recombinant Proteins
Recombination, Genetic
Salmonella - genetics
Sequence Alignment
Structure-Activity Relationship
Transcriptional Activation
Transfection
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Summary:Recent studies have demonstrated that DNA cleavage during V(D)J recombination is mediated by the RAG1 and RAG2 proteins. These proteins must therefore bind to the recombination signals, but the specific binding interaction has been difficult to study in vitro. Here, we use an in vivo one-hybrid DNA binding assay to demonstrate that RAG1, in the absence of RAG2, can mediate signal recognition via the nonamer, with the heptamer acting to enhance its binding. A region of RAG1 with sequence similarity to bacterial invertases is essential for DNA binding. Localization of RAG2 to the signal is dependent upon the presence of RAG1 and is substantially more efficient with a 12 bp spacer signal than with a 23 bp spacer signal.
ISSN:0092-8674
1097-4172
DOI:10.1016/S0092-8674(00)81343-4