EMD-7470
RAG1/2 HFC complex
EMD-7470
Single-particle3.17 Å
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Map released: 25/04/2018
Last modified: 13/03/2024
Sample Organism:
Escherichia coli K-12,
Mus musculus,
Homo sapiens
Sample: RAG1/2 in complex with nicked DNAs
Fitted models: 6cg0 (Avg. Q-score: 0.452)
Deposition Authors: Chen X
,
Kim M
Sample: RAG1/2 in complex with nicked DNAs
Fitted models: 6cg0 (Avg. Q-score: 0.452)
Deposition Authors: Chen X
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Cracking the DNA Code for V(D)J Recombination.
Kim MS,
Chuenchor W
,
Chen X
,
Cui Y,
Zhang X,
Zhou ZH,
Gellert M,
Yang W
(2018) Mol Cell , 70 , 358 - 370.e4
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(2018) Mol Cell , 70 , 358 - 370.e4
Abstract:
To initiate V(D)J recombination for generating the adaptive immune response of vertebrates, RAG1/2 recombinase cleaves DNA at a pair of recombination signal sequences, the 12- and 23-RSS. We have determined crystal and cryo-EM structures of RAG1/2 with DNA in the pre-reaction and hairpin-forming complexes up to 2.75 Å resolution. Both protein and DNA exhibit structural plasticity and undergo dramatic conformational changes. Coding-flank DNAs extensively rotate, shift, and deform for nicking and hairpin formation. Two intertwined RAG1 subunits crisscross four times between the asymmetric pair of severely bent 12/23-RSS DNAs. Location-sensitive bending of 60° and 150° in 12- and 23-RSS spacers, respectively, must occur for RAG1/2 to capture the nonamers and pair the heptamers for symmetric double-strand breakage. DNA pairing is thus sequence-context dependent and structure specific, which partly explains the "beyond 12/23" restriction. Finally, catalysis in crystallo reveals the process of DNA hairpin formation and its stabilization by interleaved base stacking.
To initiate V(D)J recombination for generating the adaptive immune response of vertebrates, RAG1/2 recombinase cleaves DNA at a pair of recombination signal sequences, the 12- and 23-RSS. We have determined crystal and cryo-EM structures of RAG1/2 with DNA in the pre-reaction and hairpin-forming complexes up to 2.75 Å resolution. Both protein and DNA exhibit structural plasticity and undergo dramatic conformational changes. Coding-flank DNAs extensively rotate, shift, and deform for nicking and hairpin formation. Two intertwined RAG1 subunits crisscross four times between the asymmetric pair of severely bent 12/23-RSS DNAs. Location-sensitive bending of 60° and 150° in 12- and 23-RSS spacers, respectively, must occur for RAG1/2 to capture the nonamers and pair the heptamers for symmetric double-strand breakage. DNA pairing is thus sequence-context dependent and structure specific, which partly explains the "beyond 12/23" restriction. Finally, catalysis in crystallo reveals the process of DNA hairpin formation and its stabilization by interleaved base stacking.