Joining-deficient RAG1 mutants block V(D)J recombination in vivo and hairpin opening in vitro.
The RAG proteins cleave at V(D)J recombination signal sequences then form a postcleavage complex with the broken ends. The role of this complex in end processing and joining, if any, is undefined. We have identified two RAG1 mutants proficient for DNA cleavage but severely defective for coding and signal joint formation, providing direct evidence that RAG1 is critical for joining in vivo and strongly suggesting that the postcleavage complex is important in end joining. We have also identified a RAG1 mutant that is severely defective for both hairpin opening in vitro and coding joint formation in vivo. These data suggest that the hairpin opening activity of the RAG proteins plays an important physiological role in V(D)J recombination.
Yarnell Schultz
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Interdisciplinary Program in Cell and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA.
Landree
M A
MA
Qiu
J X
JX
Kale
S B
SB
Roth
D B
DB
D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D020134:Catalytic Domain / Q000502:physiology; D006224:Cricetinae; D004254:DNA Nucleotidyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D018398:Homeodomain Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D066298:In Vitro Techniques; D016296:Mutagenesis / Q000235:genetics; D017434:Protein Structure, Tertiary; D011995:Recombination, Genetic / Q000502:physiology*; D045567:VDJ Recombinases
D018398:Homeodomain Proteins; C064658:RAG-1 protein; D004254:DNA Nucleotidyltransferases; D045567:VDJ Recombinases