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Yorodumi- PDB-7o80: Rabbit 80S ribosome in complex with eRF1 and ABCE1 stalled at the... -
+Open data
-Basic information
Entry | Database: PDB / ID: 7o80 | ||||||||||||
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Title | Rabbit 80S ribosome in complex with eRF1 and ABCE1 stalled at the STOP codon in the mutated SARS-CoV-2 slippery site | ||||||||||||
Components |
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Keywords | RIBOSOME / Frameshift / virus / pseudoknot | ||||||||||||
Function / homology | Function and homology information translation termination factor activity / Major pathway of rRNA processing in the nucleolus and cytosol / GTP hydrolysis and joining of the 60S ribosomal subunit / cytoplasmic translational termination / translation release factor complex / L13a-mediated translational silencing of Ceruloplasmin expression / SRP-dependent cotranslational protein targeting to membrane / Formation of a pool of free 40S subunits / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) ...translation termination factor activity / Major pathway of rRNA processing in the nucleolus and cytosol / GTP hydrolysis and joining of the 60S ribosomal subunit / cytoplasmic translational termination / translation release factor complex / L13a-mediated translational silencing of Ceruloplasmin expression / SRP-dependent cotranslational protein targeting to membrane / Formation of a pool of free 40S subunits / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / regulation of translational termination / translation release factor activity, codon specific / ribosomal subunit / sequence-specific mRNA binding / aminoacyl-tRNA hydrolase activity / regulation of G1 to G0 transition / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / exit from mitosis / laminin receptor activity / positive regulation of intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / regulation of translation involved in cellular response to UV / protein-DNA complex disassembly / positive regulation of DNA damage response, signal transduction by p53 class mediator resulting in transcription of p21 class mediator / optic nerve development / retinal ganglion cell axon guidance / mammalian oogenesis stage / G1 to G0 transition / activation-induced cell death of T cells / positive regulation of signal transduction by p53 class mediator / ubiquitin ligase inhibitor activity / phagocytic cup / ribosomal small subunit binding / 90S preribosome / TOR signaling / T cell proliferation involved in immune response / protein-RNA complex assembly / erythrocyte development / cellular response to actinomycin D / negative regulation of ubiquitin-dependent protein catabolic process / ribosomal subunit export from nucleus / ribosomal small subunit export from nucleus / translational termination / translation regulator activity / laminin binding / rough endoplasmic reticulum / gastrulation / MDM2/MDM4 family protein binding / maturation of LSU-rRNA / DNA damage response, signal transduction by p53 class mediator resulting in cell cycle arrest / cytosolic ribosome / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / class I DNA-(apurinic or apyrimidinic site) endonuclease activity / DNA-(apurinic or apyrimidinic site) lyase / rescue of stalled ribosome / ribosomal large subunit biogenesis / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / cellular response to leukemia inhibitory factor / small-subunit processome / positive regulation of translation / translational initiation / protein kinase C binding / positive regulation of apoptotic signaling pathway / positive regulation of protein-containing complex assembly / placenta development / cellular response to gamma radiation / transcription coactivator binding / mRNA 5'-UTR binding / spindle / cytoplasmic ribonucleoprotein granule / modification-dependent protein catabolic process / G1/S transition of mitotic cell cycle / protein tag activity / rRNA processing / ribosomal small subunit biogenesis / antimicrobial humoral immune response mediated by antimicrobial peptide / small ribosomal subunit rRNA binding / positive regulation of canonical Wnt signaling pathway / rhythmic process / protein guanylyltransferase activity / RNA endonuclease activity, producing 3'-phosphomonoesters / mRNA guanylyltransferase activity / 5'-3' RNA helicase activity / ribosome binding / Lyases; Phosphorus-oxygen lyases / glucose homeostasis / symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of TBK1 activity / regulation of translation / Assembly of the SARS-CoV-2 Replication-Transcription Complex (RTC) / ribosomal small subunit assembly / Maturation of replicase proteins / retina development in camera-type eye / virus receptor activity / ISG15-specific peptidase activity / Transcription of SARS-CoV-2 sgRNAs / TRAF3-dependent IRF activation pathway / Translation of Replicase and Assembly of the Replication Transcription Complex / Replication of the SARS-CoV-2 genome / snRNP Assembly / double membrane vesicle viral factory outer membrane Similarity search - Function | ||||||||||||
Biological species | Severe acute respiratory syndrome coronavirus 2 Oryctolagus cuniculus (rabbit) Bos taurus (cattle) | ||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.9 Å | ||||||||||||
Authors | Bhatt, P.R. / Scaiola, A. / Leibundgut, M.A. / Atkins, J.F. / Ban, N. | ||||||||||||
Funding support | Switzerland, Ireland, 2items
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Citation | Journal: Science / Year: 2021 Title: Structural basis of ribosomal frameshifting during translation of the SARS-CoV-2 RNA genome. Authors: Pramod R Bhatt / Alain Scaiola / Gary Loughran / Marc Leibundgut / Annika Kratzel / Romane Meurs / René Dreos / Kate M O'Connor / Angus McMillan / Jeffrey W Bode / Volker Thiel / David ...Authors: Pramod R Bhatt / Alain Scaiola / Gary Loughran / Marc Leibundgut / Annika Kratzel / Romane Meurs / René Dreos / Kate M O'Connor / Angus McMillan / Jeffrey W Bode / Volker Thiel / David Gatfield / John F Atkins / Nenad Ban / Abstract: Programmed ribosomal frameshifting is a key event during translation of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA genome that allows synthesis of the viral RNA-dependent ...Programmed ribosomal frameshifting is a key event during translation of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA genome that allows synthesis of the viral RNA-dependent RNA polymerase and downstream proteins. Here, we present the cryo-electron microscopy structure of a translating mammalian ribosome primed for frameshifting on the viral RNA. The viral RNA adopts a pseudoknot structure that lodges at the entry to the ribosomal messenger RNA (mRNA) channel to generate tension in the mRNA and promote frameshifting, whereas the nascent viral polyprotein forms distinct interactions with the ribosomal tunnel. Biochemical experiments validate the structural observations and reveal mechanistic and regulatory features that influence frameshifting efficiency. Finally, we compare compounds previously shown to reduce frameshifting with respect to their ability to inhibit SARS-CoV-2 replication, establishing coronavirus frameshifting as a target for antiviral intervention. | ||||||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | Molecule: MolmilJmol/JSmol |
-Downloads & links
-Download
PDBx/mmCIF format | 7o80.cif.gz | 5.3 MB | Display | PDBx/mmCIF format |
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PDB format | pdb7o80.ent.gz | Display | PDB format | |
PDBx/mmJSON format | 7o80.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7o80_validation.pdf.gz | 441.6 KB | Display | wwPDB validaton report |
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Full document | 7o80_full_validation.pdf.gz | 532.8 KB | Display | |
Data in XML | 7o80_validation.xml.gz | 203.5 KB | Display | |
Data in CIF | 7o80_validation.cif.gz | 336.9 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/o8/7o80 ftp://data.pdbj.org/pub/pdb/validation_reports/o8/7o80 | HTTPS FTP |
-Related structure data
Related structure data | 12758MC 7o7yC 7o7zC 7o81C M: map data used to model this data C: citing same article (ref.) |
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Similar structure data |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
-RNA chain , 7 types, 7 molecules A2AHAIATB5B7B8
#1: RNA chain | Mass: 603928.250 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Oryctolagus cuniculus (rabbit) / References: GenBank: GBCT01000564.1 |
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#9: RNA chain | Mass: 69683.008 Da / Num. of mol.: 1 / Source method: obtained synthetically Source: (synth.) Severe acute respiratory syndrome coronavirus 2 References: GenBank: NC_045512.2 |
#10: RNA chain | Mass: 15209.544 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Bos taurus (cattle) |
#11: RNA chain | Mass: 27761.496 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Bos taurus (cattle) |
#39: RNA chain | Mass: 1557519.500 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from GeneBank entry GBCN01009604.1 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: GenBank: GBCN01009604.1 |
#40: RNA chain | Mass: 38346.707 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from GeneBank entry GBCM01014045.1 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: GenBank: GBCM01014045.1 |
#41: RNA chain | Mass: 50809.047 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from GeneBank entry GBCN01009604.1 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: GenBank: GBCN01009604.1 |
-40S ribosomal protein ... , 18 types, 18 molecules AAADAZAaAcAfAgAhAkAlAnAoAqArAtAwAxAy
#2: Protein | Mass: 9480.186 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1TZ76 |
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#5: Protein | Mass: 14498.884 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1T8A2 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1T8A2 |
#12: Protein | Mass: 32970.023 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1TLT8 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1TLT8 |
#13: Protein | Mass: 30002.061 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1SS70 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1SS70 |
#15: Protein | Mass: 31146.607 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1TNM3 / Source: (natural) Oryctolagus cuniculus (rabbit) References: UniProt: G1TNM3, DNA-(apurinic or apyrimidinic site) lyase |
#18: Protein | Mass: 28751.906 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from GeneBank XP_002708126.1 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1TM55 |
#19: Protein | Mass: 22168.914 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from GeneBank NP_001272763.1 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1SVB0 |
#20: Protein | Mass: 24263.387 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from GeneBank entry XP_017201844.1 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1TJW1 |
#23: Protein | Mass: 18468.826 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1TRM4 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1TRM4 |
#24: Protein | Mass: 14538.987 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1SFR8 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1SFR8 |
#26: Protein | Mass: 16302.772 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1T1F0 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1T1F0 |
#27: Protein | Mass: 17049.182 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1U0Q2 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1U0Q2 |
#29: Protein | Mass: 15552.119 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1TU13 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1TU13 |
#30: Protein | Mass: 17801.814 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1TPG3 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1TPG3 |
#32: Protein | Mass: 13398.763 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1SIZ2 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1SIZ2 |
#35: Protein | Mass: 15860.666 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1SZ47 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1SZ47 |
#36: Protein | Mass: 15107.924 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry A0A5F9CBF4 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1T3D8 |
#37: Protein | Mass: 13645.028 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1TDB3 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1TDB3 |
+Ribosomal protein ... , 29 types, 29 molecules ABACAEAbAdAeAiAjApAsAuAvBABBBFBGBJBLBMBNBOBQBSBUBVBYBjBrBt
+Protein , 22 types, 22 molecules AFAGAmBDBKBPBTBWBXBcBdBeBfBhBkBlBoBpBsBvByBz
-60S ribosomal protein ... , 12 types, 12 molecules AzBCBEBHBIBRBZBaBbBgBiBm
#38: Protein/peptide | Mass: 3473.451 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry A0A087WNH4 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: A0A087WNH4 |
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#44: Protein | Mass: 46430.895 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1SVW5 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1SVW5 |
#46: Protein | Mass: 33028.336 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from GeneBank entry XP_017205046.1 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1SKF7 |
#49: Protein | Mass: 21871.418 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1SWI6 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1SWI6 |
#50: Protein | Mass: 24643.057 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry B7NZQ2 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: B7NZQ2 |
#59: Protein | Mass: 23535.281 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from GeneBank entry DN889037.1 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: Q3T0W9 |
#67: Protein | Mass: 15835.831 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1TXF6 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1TXF6 |
#68: Protein | Mass: 16635.551 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1SNY0 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1SNY0 |
#69: Protein | Mass: 26721.727 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1SGR6 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1SGR6 |
#74: Protein | Mass: 13326.074 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from UniProt entry G1TXG5 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1TXG5 |
#76: Protein | Mass: 12263.834 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from GeneBank entry XP_002708713.1 / Source: (natural) Oryctolagus cuniculus (rabbit) / References: UniProt: G1TTQ5 |
#80: Protein | Mass: 14800.474 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Sequence from GeneBank entry DN885652.1 / Source: (natural) Oryctolagus cuniculus (rabbit) |
-Non-polymers , 7 types, 2478 molecules
#89: Chemical | ChemComp-SPD / | ||||||||||
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#90: Chemical | ChemComp-MG / #91: Chemical | ChemComp-UNX / #92: Chemical | ChemComp-ZN / #93: Chemical | ChemComp-GTP / | #94: Chemical | #95: Water | ChemComp-HOH / | |
-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component |
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Molecular weight | Experimental value: NO | ||||||||||||||||||||||||||||||||||||||||||
Source (natural) |
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Source (recombinant) |
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Buffer solution | pH: 7.4 | ||||||||||||||||||||||||||||||||||||||||||
Specimen | Conc.: 0.3 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | ||||||||||||||||||||||||||||||||||||||||||
Specimen support | Grid material: COPPER / Grid mesh size: 400 divisions/in. / Grid type: Quantifoil R2/2 | ||||||||||||||||||||||||||||||||||||||||||
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE-PROPANE / Humidity: 100 % / Chamber temperature: 277 K |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 81000 X / Calibrated magnification: 56604 X / Cs: 2.7 mm / C2 aperture diameter: 100 µm |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Electron dose: 60 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
EM imaging optics | Energyfilter name: GIF Quantum LS / Energyfilter slit width: 20 eV |
-Processing
EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||
3D reconstruction | Resolution: 2.9 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 55057 / Symmetry type: POINT |