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Yorodumi- EMDB-10209: Cryo-EM structure of the Type III-B Cmr-beta bound to cognate tar... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-10209 | ||||||||||||
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Title | Cryo-EM structure of the Type III-B Cmr-beta bound to cognate target RNA | ||||||||||||
Map data | |||||||||||||
Sample |
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Keywords | CRISPR-Cas / Effector complex / nuclease / cyclic oligo-adenylate synthase / ANTIVIRAL PROTEIN | ||||||||||||
Function / homology | Function and homology information | ||||||||||||
Biological species | Sulfolobus islandicus REY15A (acidophilic) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.52 Å | ||||||||||||
Authors | Sofos N / Montoya G | ||||||||||||
Funding support | Denmark, 3 items
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Citation | Journal: Mol Cell / Year: 2020 Title: Structures of the Cmr-β Complex Reveal the Regulation of the Immunity Mechanism of Type III-B CRISPR-Cas. Authors: Nicholas Sofos / Mingxia Feng / Stefano Stella / Tillmann Pape / Anders Fuglsang / Jinzhong Lin / Qihong Huang / Yingjun Li / Qunxin She / Guillermo Montoya / Abstract: Cmr-β is a type III-B CRISPR-Cas complex that, upon target RNA recognition, unleashes a multifaceted immune response against invading genetic elements, including single-stranded DNA (ssDNA) ...Cmr-β is a type III-B CRISPR-Cas complex that, upon target RNA recognition, unleashes a multifaceted immune response against invading genetic elements, including single-stranded DNA (ssDNA) cleavage, cyclic oligoadenylate synthesis, and also a unique UA-specific single-stranded RNA (ssRNA) hydrolysis by the Cmr2 subunit. Here, we present the structure-function relationship of Cmr-β, unveiling how binding of the target RNA regulates the Cmr2 activities. Cryoelectron microscopy (cryo-EM) analysis revealed the unique subunit architecture of Cmr-β and captured the complex in different conformational stages of the immune response, including the non-cognate and cognate target-RNA-bound complexes. The binding of the target RNA induces a conformational change of Cmr2, which together with the complementation between the 5' tag in the CRISPR RNAs (crRNA) and the 3' antitag of the target RNA activate different configurations in a unique loop of the Cmr3 subunit, which acts as an allosteric sensor signaling the self- versus non-self-recognition. These findings highlight the diverse defense strategies of type III complexes. | ||||||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_10209.map.gz | 391.3 MB | EMDB map data format | |
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Header (meta data) | emd-10209-v30.xml emd-10209.xml | 32.3 KB 32.3 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_10209_fsc.xml | 16.3 KB | Display | FSC data file |
Images | emd_10209.png | 19.4 KB | ||
Filedesc metadata | emd-10209.cif.gz | 8.6 KB | ||
Others | emd_10209_half_map_1.map.gz emd_10209_half_map_2.map.gz | 63.1 MB 63.1 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-10209 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-10209 | HTTPS FTP |
-Validation report
Summary document | emd_10209_validation.pdf.gz | 908.6 KB | Display | EMDB validaton report |
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Full document | emd_10209_full_validation.pdf.gz | 908.1 KB | Display | |
Data in XML | emd_10209_validation.xml.gz | 23.3 KB | Display | |
Data in CIF | emd_10209_validation.cif.gz | 31.4 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10209 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10209 | HTTPS FTP |
-Related structure data
Related structure data | 6sicMC 6s6bC 6s8bC 6s8eC 6s91C 6sh8C 6shbC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_10209.map.gz / Format: CCP4 / Size: 421.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Half map: #2
File | emd_10209_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_10209_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Type III-B Cmr-beta binary complex, crRNA-bound
+Supramolecule #1: Type III-B Cmr-beta binary complex, crRNA-bound
+Supramolecule #2: Type III-B Cmr-beta binary complex, crRNA-bound
+Supramolecule #3: Type III-B Cmr-beta binary complex, crRNA-bound
+Supramolecule #4: Type III-B Cmr-beta binary complex, crRNA-bound
+Macromolecule #1: CRISPR-associated protein, Cmr5 family
+Macromolecule #2: CRISPR-associated RAMP protein, Cmr4 family
+Macromolecule #3: CRISPR-associated protein, Cmr3 family
+Macromolecule #4: CRISPR-associated RAMP protein, Cmr6 family
+Macromolecule #5: CRISPR-associated RAMP protein, Cmr1 family
+Macromolecule #6: CRISPR-associated protein, Cmr2 family
+Macromolecule #7: CRISPR-associated protein Cmrx
+Macromolecule #8: Cognate target RNA
+Macromolecule #9: crRNA
+Macromolecule #10: ZINC ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.8 mg/mL | |||||||||||||||
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Buffer | pH: 6 Component:
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Grid | Model: Quantifoil, UltrAuFoil / Material: GOLD / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 60 sec. / Pretreatment - Atmosphere: AIR / Details: 5 mA (Leica EM ACE200) | |||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV / Details: 3-4 s blotting before plunging. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: COUNTING / Number grids imaged: 1 / Number real images: 3785 / Average exposure time: 40.0 sec. / Average electron dose: 40.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.1 mm / Nominal defocus max: 2.6 µm / Nominal defocus min: 1.7 µm / Nominal magnification: 96000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |