+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-22413 | |||||||||
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Title | 1:1 cGAS(mask)-NCP map | |||||||||
Map data | 1:! cGAS(mask)-nucleosome map | |||||||||
Sample |
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Method | single particle reconstruction / cryo EM / Resolution: 3.9 Å | |||||||||
Authors | Boyer JA / Spangler CJ / Strauss JD / McGinty RK / Zhang Q | |||||||||
Funding support | United States, 2 items
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Citation | Journal: Science / Year: 2020 Title: Structural basis of nucleosome-dependent cGAS inhibition. Authors: Joshua A Boyer / Cathy J Spangler / Joshua D Strauss / Andrew P Cesmat / Pengda Liu / Robert K McGinty / Qi Zhang / Abstract: Cyclic guanosine monophosphate (GMP)-adenosine monophosphate (AMP) synthase (cGAS) recognizes cytosolic foreign or damaged DNA to activate the innate immune response to infection, inflammatory ...Cyclic guanosine monophosphate (GMP)-adenosine monophosphate (AMP) synthase (cGAS) recognizes cytosolic foreign or damaged DNA to activate the innate immune response to infection, inflammatory diseases, and cancer. By contrast, cGAS reactivity against self-DNA in the nucleus is suppressed by chromatin tethering. We report a 3.3-angstrom-resolution cryo-electron microscopy structure of cGAS in complex with the nucleosome core particle. The structure reveals that cGAS uses two conserved arginines to anchor to the nucleosome acidic patch. The nucleosome-binding interface exclusively occupies the strong double-stranded DNA (dsDNA)-binding surface on cGAS and sterically prevents cGAS from oligomerizing into the functionally active 2:2 cGAS-dsDNA state. These findings provide a structural basis for how cGAS maintains an inhibited state in the nucleus and further exemplify the role of the nucleosome in regulating diverse nuclear protein functions. | |||||||||
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_22413.map.gz | 9 MB | EMDB map data format | |
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Header (meta data) | emd-22413-v30.xml emd-22413.xml | 15.2 KB 15.2 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_22413_fsc.xml | 11.6 KB | Display | FSC data file |
Images | emd_22413.png | 79.1 KB | ||
Others | emd_22413_additional.map.gz emd_22413_additional_1.map.gz | 73.9 MB 73.9 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-22413 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-22413 | HTTPS FTP |
-Validation report
Summary document | emd_22413_validation.pdf.gz | 79 KB | Display | EMDB validaton report |
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Full document | emd_22413_full_validation.pdf.gz | 78.1 KB | Display | |
Data in XML | emd_22413_validation.xml.gz | 493 B | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-22413 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-22413 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_22413.map.gz / Format: CCP4 / Size: 137.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | 1:! cGAS(mask)-nucleosome map | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.91 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: 1:1 cGAS-nucleosome composite map
File | emd_22413_additional.map | ||||||||||||
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Annotation | 1:1 cGAS-nucleosome composite map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: 1:1 cGAS-nucleosome composite map
File | emd_22413_additional_1.map | ||||||||||||
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Annotation | 1:1 cGAS-nucleosome composite map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : 1:1 cGAS-nucleosome complex
Entire | Name: 1:1 cGAS-nucleosome complex |
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Components |
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-Supramolecule #1: 1:1 cGAS-nucleosome complex
Supramolecule | Name: 1:1 cGAS-nucleosome complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#7 / Details: mouse cGAS bound to the nucleosome in a 1:1 ratio |
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Molecular weight | Theoretical: 250 KDa |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 1 mg/mL | ||||||||||||
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Buffer | pH: 7.5 Component:
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Grid | Model: Quantifoil / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Details: Instrument: Pelco easiGlow | ||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 293 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Number real images: 2100 / Average electron dose: 53.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |