5ZE1

Hairpin Forming Complex, RAG1/2-Nicked 12RSS/23RSS complex in 2mM Mn2+ for 10 min at 4'C

Summary for 5ZE1

• Entry DOI: 10.2210/pdb5ze1/pdb
• Related: 4WWX 5EZ2 5ZDZ 5ZE0 6CG0 6CIJ 6CIK 6CIL 6CIM
• Descriptor: mouse RAG1, ZINC ION, MANGANESE (II) ION, ... (14 entities in total)
• Functional Keywords: v(d)j recombination, rag1-2-12rss-23rss complex, hairpin forming complex, dna binding protein
• Biological source: Mus musculus (Mouse); More
• Cellular location: Nucleus : P15919 P21784 P63158
• Total number of polymer chains: 11
• Total formula weight: 311097.42

Authors

Kim, M.S.,Chuenchor, W.,Chen, X.,Gellert, M.,Yang, W. (deposition date: 2018-02-25, release date: 2018-04-25, Last modification date: 2024-03-27)

Primary citation

Kim, M.S.,Chuenchor, W.,Chen, X.,Cui, Y.,Zhang, X.,Zhou, Z.H.,Gellert, M.,Yang, W.
Cracking the DNA Code for V(D)J Recombination
Mol. Cell, 70:358-370.e4, 2018

PubMed 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.

PubMed: 29628308
DOI: 10.1016/j.molcel.2018.03.008

Experimental method

X-RAY DIFFRACTION (3 Å)