EMD-7470

Single-particle
3.17 Å
EMD-7470 Deposition: 19/02/2018
Map released: 25/04/2018
Last modified: 13/03/2024
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links

EMD-7470

RAG1/2 HFC complex

EMD-7470

Single-particle
3.17 Å
EMD-7470 Deposition: 19/02/2018
Map released: 25/04/2018
Last modified: 13/03/2024
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links
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
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
PUBMED: 29628308
DOI: doi:10.1016/j.molcel.2018.03.008
ISSN: 1097-4164
ASTM: MOCEFL
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.