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TitleMolecular Mechanism of V(D)J Recombination from Synaptic RAG1-RAG2 Complex Structures.
Journal, issue, pagesCell, Vol. 163, Issue 5, Page 1138-1152, Year 2015
Publish dateNov 19, 2015
AuthorsHeng Ru / Melissa G Chambers / Tian-Min Fu / Alexander B Tong / Maofu Liao / Hao Wu /
PubMed AbstractDiverse repertoires of antigen-receptor genes that result from combinatorial splicing of coding segments by V(D)J recombination are hallmarks of vertebrate immunity. The (RAG1-RAG2)2 recombinase (RAG) ...Diverse repertoires of antigen-receptor genes that result from combinatorial splicing of coding segments by V(D)J recombination are hallmarks of vertebrate immunity. The (RAG1-RAG2)2 recombinase (RAG) recognizes recombination signal sequences (RSSs) containing a heptamer, a spacer of 12 or 23 base pairs, and a nonamer (12-RSS or 23-RSS) and introduces precise breaks at RSS-coding segment junctions. RAG forms synaptic complexes only with one 12-RSS and one 23-RSS, a dogma known as the 12/23 rule that governs the recombination fidelity. We report cryo-electron microscopy structures of synaptic RAG complexes at up to 3.4 Å resolution, which reveal a closed conformation with base flipping and base-specific recognition of RSSs. Distortion at RSS-coding segment junctions and base flipping in coding segments uncover the two-metal-ion catalytic mechanism. Induced asymmetry involving tilting of the nonamer-binding domain dimer of RAG1 upon binding of HMGB1-bent 12-RSS or 23-RSS underlies the molecular mechanism for the 12/23 rule.
External linksCell / PubMed:26548953 / PubMed Central
MethodsEM (single particle)
Resolution3.3 - 9.0 Å
Structure data

6487

3jbx

EMDB-6487: Cryo-electron microscopy structure of RAG SEC (C2 symmetry)
PDB-3jbx: Cryo-electron microscopy structure of RAG Signal End Complex (C2 symmetry)
Method: EM (single particle) / Resolution: 3.4 Å

6488

3jby

EMDB-6488: Cryo-electron microscopy structure of RAG PC (C2 symmetry)
PDB-3jby: Cryo-electron microscopy structure of RAG Paired Complex (C2 symmetry)
Method: EM (single particle) / Resolution: 3.7 Å

6489

3jbw

EMDB-6489: Cryo-electron microscopy structure of RAG PC (with NBD, no symmetry)
PDB-3jbw: Cryo-electron microscopy structure of RAG Paired Complex (with NBD, no symmetry)
Method: EM (single particle) / Resolution: 4.6 Å

EMDB-6490:
Cryo-electron microscopy structure of Apo-RAG (C2 symmetry)
Method: EM (single particle) / Resolution: 9.0 Å

EMDB-6491:
Cryo-electron microscopy structure from the combined data of RAG SEC and PC (C2 symmetry)
Method: EM (single particle) / Resolution: 3.3 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

ChemComp-CA:
Unknown entry

Source
  • danio rerio (zebrafish)
  • homo sapiens (human)
KeywordsRECOMBINATION/DNA / RAG1 / RAG2 / V(D)J recombination / Paired complex / Antigen receptor gene recombination / T and B cell development / RECOMBINATION-DNA complex / Signal end complex

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