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 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.
Difilippantonio
M J
MJ
Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06520-8011, USA.
McMahan
C J
CJ
Eastman
Q M
QM
Spanopoulou
E
E
Schatz
D G
DG
D000595:Amino Acid Sequence; D002460:Cell Line; D004254:DNA Nucleotidyltransferases / Q000235:genetics; D004268:DNA-Binding Proteins / Q000502:physiology; D005803:Genes, Immunoglobulin; D018398:Homeodomain Proteins; D006801:Humans; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D011485:Protein Binding; D011506:Proteins / Q000502:physiology*; D011994:Recombinant Proteins; D011995:Recombination, Genetic; D012475:Salmonella / Q000235:genetics; D016415:Sequence Alignment; D013329:Structure-Activity Relationship; D015533:Transcriptional Activation; D014162:Transfection
D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D046911:Macromolecular Substances; D009687:Nuclear Proteins; D011506:Proteins; C499311:RAG2 protein, human; D011994:Recombinant Proteins; C064655:V(D)J recombination activating protein 2; C064658:RAG-1 protein; D004254:DNA Nucleotidyltransferases; C059997:Hin recombinase