+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-11276 | ||||||||||||
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Title | SARS-CoV-2 Nsp1 bound to the human 40S ribosomal subunit | ||||||||||||
Map data | In vitro reconstituted Nsp1-40S complex. Focused refined map of 40S head. | ||||||||||||
Sample |
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Keywords | Translational Inhibition / SARS-CoV-2 / Immune Evasion / Human Ribosome / VIRAL PROTEIN | ||||||||||||
Function / homology | Function and homology information : / negative regulation of endoplasmic reticulum unfolded protein response / oxidized pyrimidine DNA binding / response to TNF agonist / positive regulation of base-excision repair / protein tyrosine kinase inhibitor activity / positive regulation of respiratory burst involved in inflammatory response / positive regulation of intrinsic apoptotic signaling pathway in response to DNA damage / positive regulation of gastrulation / nucleolus organization ...: / negative regulation of endoplasmic reticulum unfolded protein response / oxidized pyrimidine DNA binding / response to TNF agonist / positive regulation of base-excision repair / protein tyrosine kinase inhibitor activity / positive regulation of respiratory burst involved in inflammatory response / positive regulation of intrinsic apoptotic signaling pathway in response to DNA damage / positive regulation of gastrulation / nucleolus organization / regulation of adenylate cyclase-activating G protein-coupled receptor signaling pathway / IRE1-RACK1-PP2A complex / positive regulation of endodeoxyribonuclease activity / positive regulation of Golgi to plasma membrane protein transport / TNFR1-mediated ceramide production / negative regulation of RNA splicing / negative regulation of DNA repair / negative regulation of intrinsic apoptotic signaling pathway in response to hydrogen peroxide / supercoiled DNA binding / oxidized purine DNA binding / neural crest cell differentiation / NF-kappaB complex / ubiquitin-like protein conjugating enzyme binding / regulation of establishment of cell polarity / positive regulation of ubiquitin-protein transferase activity / negative regulation of phagocytosis / rRNA modification in the nucleus and cytosol / erythrocyte homeostasis / Formation of the ternary complex, and subsequently, the 43S complex / cytoplasmic side of rough endoplasmic reticulum membrane / laminin receptor activity / pigmentation / protein kinase A binding / negative regulation of ubiquitin protein ligase activity / Ribosomal scanning and start codon recognition / ion channel inhibitor activity / Translation initiation complex formation / mammalian oogenesis stage / fibroblast growth factor binding / positive regulation of mitochondrial depolarization / activation-induced cell death of T cells / positive regulation of T cell receptor signaling pathway / negative regulation of peptidyl-serine phosphorylation / iron-sulfur cluster binding / negative regulation of Wnt signaling pathway / positive regulation of activated T cell proliferation / monocyte chemotaxis / Protein hydroxylation / regulation of cell division / BH3 domain binding / cysteine-type endopeptidase activator activity involved in apoptotic process / mTORC1-mediated signalling / SARS-CoV-1 modulates host translation machinery / Peptide chain elongation / positive regulation of intrinsic apoptotic signaling pathway by p53 class mediator / Selenocysteine synthesis / positive regulation of signal transduction by p53 class mediator / Formation of a pool of free 40S subunits / ubiquitin ligase inhibitor activity / Eukaryotic Translation Termination / phagocytic cup / negative regulation of respiratory burst involved in inflammatory response / Response of EIF2AK4 (GCN2) to amino acid deficiency / SRP-dependent cotranslational protein targeting to membrane / Viral mRNA Translation / negative regulation of phosphatidylinositol 3-kinase/protein kinase B signal transduction / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / GTP hydrolysis and joining of the 60S ribosomal subunit / L13a-mediated translational silencing of Ceruloplasmin expression / TOR signaling / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / T cell proliferation involved in immune response / regulation of translational fidelity / spindle assembly / positive regulation of cell cycle / Major pathway of rRNA processing in the nucleolus and cytosol / erythrocyte development / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / negative regulation of ubiquitin-dependent protein catabolic process / Amplification of signal from unattached kinetochores via a MAD2 inhibitory signal / Protein methylation / positive regulation of intrinsic apoptotic signaling pathway / ribosomal small subunit export from nucleus / Nuclear events stimulated by ALK signaling in cancer / translation regulator activity / signaling adaptor activity / laminin binding / negative regulation of smoothened signaling pathway / stress granule assembly / Mitotic Prometaphase / rough endoplasmic reticulum / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / antiviral innate immune response / positive regulation of JUN kinase activity / EML4 and NUDC in mitotic spindle formation / gastrulation / MDM2/MDM4 family protein binding / Maturation of protein E / Maturation of protein E / ER Quality Control Compartment (ERQC) Similarity search - Function | ||||||||||||
Biological species | Homo sapiens (human) / Severe acute respiratory syndrome coronavirus 2 | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.6 Å | ||||||||||||
Authors | Thoms M / Buschauer R | ||||||||||||
Funding support | Germany, 3 items
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Citation | Journal: Science / Year: 2020 Title: Structural basis for translational shutdown and immune evasion by the Nsp1 protein of SARS-CoV-2. Authors: Matthias Thoms / Robert Buschauer / Michael Ameismeier / Lennart Koepke / Timo Denk / Maximilian Hirschenberger / Hanna Kratzat / Manuel Hayn / Timur Mackens-Kiani / Jingdong Cheng / Jan H ...Authors: Matthias Thoms / Robert Buschauer / Michael Ameismeier / Lennart Koepke / Timo Denk / Maximilian Hirschenberger / Hanna Kratzat / Manuel Hayn / Timur Mackens-Kiani / Jingdong Cheng / Jan H Straub / Christina M Stürzel / Thomas Fröhlich / Otto Berninghausen / Thomas Becker / Frank Kirchhoff / Konstantin M J Sparrer / Roland Beckmann / Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the current coronavirus disease 2019 (COVID-19) pandemic. A major virulence factor of SARS-CoVs is the ...Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the current coronavirus disease 2019 (COVID-19) pandemic. A major virulence factor of SARS-CoVs is the nonstructural protein 1 (Nsp1), which suppresses host gene expression by ribosome association. Here, we show that Nsp1 from SARS-CoV-2 binds to the 40 ribosomal subunit, resulting in shutdown of messenger RNA (mRNA) translation both in vitro and in cells. Structural analysis by cryo-electron microscopy of in vitro-reconstituted Nsp1-40 and various native Nsp1-40 and -80 complexes revealed that the Nsp1 C terminus binds to and obstructs the mRNA entry tunnel. Thereby, Nsp1 effectively blocks retinoic acid-inducible gene I-dependent innate immune responses that would otherwise facilitate clearance of the infection. Thus, the structural characterization of the inhibitory mechanism of Nsp1 may aid structure-based drug design against SARS-CoV-2. | ||||||||||||
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_11276.map.gz | 164.3 MB | EMDB map data format | |
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Header (meta data) | emd-11276-v30.xml emd-11276.xml | 58.8 KB 58.8 KB | Display Display | EMDB header |
Images | emd_11276.png | 49.4 KB | ||
Filedesc metadata | emd-11276.cif.gz | 11.6 KB | ||
Others | emd_11276_additional_1.map.gz emd_11276_additional_2.map.gz emd_11276_additional_3.map.gz | 8.1 MB 2.4 MB 96.8 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-11276 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-11276 | HTTPS FTP |
-Validation report
Summary document | emd_11276_validation.pdf.gz | 387.8 KB | Display | EMDB validaton report |
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Full document | emd_11276_full_validation.pdf.gz | 387.3 KB | Display | |
Data in XML | emd_11276_validation.xml.gz | 7.5 KB | Display | |
Data in CIF | emd_11276_validation.cif.gz | 8.5 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-11276 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-11276 | HTTPS FTP |
-Related structure data
Related structure data | 6zlwMC 6zm7C 6zmeC 6zmiC 6zmoC 6zmtC 6zn5C 6zonC 6zp4C 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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Related items in Molecule of the Month |
-Map
File | Download / File: emd_11276.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | In vitro reconstituted Nsp1-40S complex. Focused refined map of 40S head. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.059 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: In vitro reconstituted Nsp1-40S complex. Focused refined map...
File | emd_11276_additional_1.map | ||||||||||||
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Annotation | In vitro reconstituted Nsp1-40S complex. Focused refined map of 40S body. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Native Nsp1-40S complex.
File | emd_11276_additional_2.map | ||||||||||||
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Annotation | Native Nsp1-40S complex. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Native Nsp1-40S complex.
File | emd_11276_additional_3.map | ||||||||||||
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Annotation | Native Nsp1-40S complex. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : SARS-CoV-2 Nsp1 bound to the human 40S ribosomal subunit
+Supramolecule #1: SARS-CoV-2 Nsp1 bound to the human 40S ribosomal subunit
+Supramolecule #2: Human 40S ribosomal subunit
+Supramolecule #3: SARS-CoV-2 Nsp1
+Macromolecule #1: 40S ribosomal protein SA
+Macromolecule #2: 40S ribosomal protein S3a
+Macromolecule #3: 40S ribosomal protein S2
+Macromolecule #4: 40S ribosomal protein S4, X isoform
+Macromolecule #5: 40S ribosomal protein S3
+Macromolecule #6: 40S ribosomal protein S6
+Macromolecule #7: 40S ribosomal protein S7
+Macromolecule #8: 40S ribosomal protein S8
+Macromolecule #9: 40S ribosomal protein S9
+Macromolecule #10: 40S ribosomal protein S5
+Macromolecule #11: 40S ribosomal protein S11
+Macromolecule #12: 40S ribosomal protein S10
+Macromolecule #13: 40S ribosomal protein S13
+Macromolecule #14: 40S ribosomal protein S12
+Macromolecule #15: 40S ribosomal protein S14
+Macromolecule #16: 40S ribosomal protein S15
+Macromolecule #17: 40S ribosomal protein S16
+Macromolecule #18: 40S ribosomal protein S17
+Macromolecule #19: 40S ribosomal protein S18
+Macromolecule #20: 40S ribosomal protein S19
+Macromolecule #21: 40S ribosomal protein S20
+Macromolecule #22: 40S ribosomal protein S15a
+Macromolecule #23: 40S ribosomal protein S23
+Macromolecule #24: 40S ribosomal protein S24
+Macromolecule #25: 40S ribosomal protein S21
+Macromolecule #26: 40S ribosomal protein S25
+Macromolecule #27: 40S ribosomal protein S27
+Macromolecule #28: 40S ribosomal protein S26
+Macromolecule #29: 40S ribosomal protein S28
+Macromolecule #30: 40S ribosomal protein S30
+Macromolecule #31: 40S ribosomal protein S29
+Macromolecule #32: Ubiquitin-40S ribosomal protein S27a
+Macromolecule #33: 60S ribosomal protein L41
+Macromolecule #34: Receptor of activated protein C kinase 1
+Macromolecule #36: Non-structural protein 1
+Macromolecule #35: 18S ribosomal RNA
+Macromolecule #37: ZINC ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
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Vitrification | Cryogen name: ETHANE |
-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: 44.8 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Startup model | Type of model: OTHER |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 2.6 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 173060 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |