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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-20035 | |||||||||
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Title | Cryo-EM structure of mouse RAG1/2 NFC complex (DNA2) | |||||||||
![]() | mouse RAG1/2 NFC complex | |||||||||
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![]() | V(D)J recombination / DNA Transposition / RAG / SCID / RECOMBINATION / RECOMBINATION-DNA complex | |||||||||
Function / homology | ![]() non-sequence-specific DNA binding, bending / DNA-binding transcription factor binding => GO:0140297 / regulation of restriction endodeoxyribonuclease activity / heterochromatin formation => GO:0031507 / regulation of tolerance induction / positive regulation of mismatch repair / negative regulation of apoptotic cell clearance / negative regulation of RNA polymerase II transcription preinitiation complex assembly / DNA geometric change / mature B cell differentiation involved in immune response ...non-sequence-specific DNA binding, bending / DNA-binding transcription factor binding => GO:0140297 / regulation of restriction endodeoxyribonuclease activity / heterochromatin formation => GO:0031507 / regulation of tolerance induction / positive regulation of mismatch repair / negative regulation of apoptotic cell clearance / negative regulation of RNA polymerase II transcription preinitiation complex assembly / DNA geometric change / mature B cell differentiation involved in immune response / T-helper 1 cell activation / C-X-C chemokine binding / T-helper 1 cell differentiation / positive regulation of dendritic cell differentiation / DNA recombinase complex / negative regulation of CD4-positive, alpha-beta T cell differentiation / B cell homeostatic proliferation / DN2 thymocyte differentiation / negative regulation of T cell differentiation in thymus / endodeoxyribonuclease complex / neutrophil clearance / pre-B cell allelic exclusion / positive regulation of interleukin-1 production / positive regulation of organ growth / regulation of behavioral fear response / alphav-beta3 integrin-HMGB1 complex / bubble DNA binding / V(D)J recombination / negative regulation of T cell apoptotic process / phosphatidylinositol-3,4-bisphosphate binding / inflammatory response to antigenic stimulus / negative regulation of thymocyte apoptotic process / positive regulation of chemokine (C-X-C motif) ligand 2 production / positive regulation of monocyte chemotaxis / supercoiled DNA binding / phosphatidylinositol-3,5-bisphosphate binding / DNA binding, bending / positive regulation of T cell differentiation / positive regulation of vascular endothelial cell proliferation / regulation of T cell differentiation / organ growth / T cell lineage commitment / positive regulation of DNA binding / B cell lineage commitment / positive regulation of activated T cell proliferation / phosphatidylserine binding / positive regulation of cysteine-type endopeptidase activity involved in apoptotic process / T cell homeostasis / phosphatidylinositol-3,4,5-trisphosphate binding / positive regulation of interleukin-10 production / negative regulation of blood vessel endothelial cell migration / endoplasmic reticulum-Golgi intermediate compartment / negative regulation of type II interferon production / T cell differentiation / positive regulation of blood vessel endothelial cell migration / protein autoubiquitination / positive regulation of autophagy / DNA polymerase binding / four-way junction DNA binding / condensed chromosome / phosphatidylinositol-4,5-bisphosphate binding / activation of innate immune response / positive regulation of interleukin-12 production / methylated histone binding / transcription repressor complex / phosphatidylinositol binding / B cell differentiation / thymus development / positive regulation of interleukin-8 production / lipopolysaccharide binding / positive regulation of JNK cascade / visual learning / RING-type E3 ubiquitin transferase / autophagy / positive regulation of interleukin-6 production / ubiquitin-protein transferase activity / chemotaxis / ubiquitin protein ligase activity / positive regulation of tumor necrosis factor production / integrin binding / chromatin organization / histone binding / positive regulation of cytosolic calcium ion concentration / T cell differentiation in thymus / endonuclease activity / DNA recombination / adaptive immune response / sequence-specific DNA binding / damaged DNA binding / transcription coactivator activity / Hydrolases; Acting on ester bonds / positive regulation of ERK1 and ERK2 cascade / transcription cis-regulatory region binding / lyase activity / endosome / defense response to bacterium / positive regulation of apoptotic process / innate immune response / DNA repair / chromatin binding Similarity search - Function | |||||||||
Biological species | ![]() ![]() ![]() ![]() ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.29 Å | |||||||||
![]() | Chen X / Cui Y / Zhou ZH / Yang W / Gellert M | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Cutting antiparallel DNA strands in a single active site. Authors: Xuemin Chen / Yanxiang Cui / Robert B Best / Huaibin Wang / Z Hong Zhou / Wei Yang / Martin Gellert / ![]() Abstract: A single enzyme active site that catalyzes multiple reactions is a well-established biochemical theme, but how one nuclease site cleaves both DNA strands of a double helix has not been well ...A single enzyme active site that catalyzes multiple reactions is a well-established biochemical theme, but how one nuclease site cleaves both DNA strands of a double helix has not been well understood. In analyzing site-specific DNA cleavage by the mammalian RAG1-RAG2 recombinase, which initiates V(D)J recombination, we find that the active site is reconfigured for the two consecutive reactions and the DNA double helix adopts drastically different structures. For initial nicking of the DNA, a locally unwound and unpaired DNA duplex forms a zipper via alternating interstrand base stacking, rather than melting as generally thought. The second strand cleavage and formation of a hairpin-DNA product requires a global scissor-like movement of protein and DNA, delivering the scissile phosphate into the rearranged active site. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 78.3 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 17.9 KB 17.9 KB | Display Display | ![]() |
Images | ![]() | 59.5 KB | ||
Filedesc metadata | ![]() | 7.1 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6oerMC ![]() 6oemC ![]() 6oenC ![]() 6oeoC ![]() 6oepC ![]() 6oeqC ![]() 6v0vC C: citing same article ( M: atomic model generated by this map |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | mouse RAG1/2 NFC complex | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.07 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
+Entire : RAG1/2 Nick-forming complex (DNA2)
+Supramolecule #1: RAG1/2 Nick-forming complex (DNA2)
+Macromolecule #1: V(D)J recombination-activating protein 1
+Macromolecule #2: V(D)J recombination-activating protein 2
+Macromolecule #7: High mobility group protein B1
+Macromolecule #3: DNA (46-MER)
+Macromolecule #4: DNA (46-MER)
+Macromolecule #5: DNA (57-MER)
+Macromolecule #6: DNA (57-MER)
+Macromolecule #8: ZINC ION
+Macromolecule #9: CALCIUM ION
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.4 |
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Grid | Details: unspecified |
Vitrification | Cryogen name: ETHANE |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Startup model | Type of model: NONE |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.29 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 333280 |
Initial angle assignment | Type: NOT APPLICABLE |
Final angle assignment | Type: NOT APPLICABLE |