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- EMDB-14367: Tomogram showing SARS-CoV-2 virions within a viral containing com... -

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Entry
Database: EMDB / ID: EMD-14367
TitleTomogram showing SARS-CoV-2 virions within a viral containing compartment of an infected airway cell
Map dataTomogram showing SARS-CoV-2 virions within a viral containing compartment of an infected airway cell
Sample
  • Cell: Airway epithelial cells infected with SARS-CoV-2
Biological speciesSevere acute respiratory syndrome coronavirus 2
Methodelectron tomography / negative staining
AuthorsPinto AL / Rai RK / Brown JC / Griffin P / Edgar JR / Shah A / Singanayagam A / Hogg C / Barclay WS / Futter CE / Burgoyne T
Funding support1 items
OrganizationGrant numberCountry
Not funded
CitationJournal: Nat Commun / Year: 2022
Title: Ultrastructural insight into SARS-CoV-2 entry and budding in human airway epithelium.
Authors: Andreia L Pinto / Ranjit K Rai / Jonathan C Brown / Paul Griffin / James R Edgar / Anand Shah / Aran Singanayagam / Claire Hogg / Wendy S Barclay / Clare E Futter / Thomas Burgoyne /
Abstract: Ultrastructural studies of SARS-CoV-2 infected cells are crucial to better understand the mechanisms of viral entry and budding within host cells. Here, we examined human airway epithelium infected ...Ultrastructural studies of SARS-CoV-2 infected cells are crucial to better understand the mechanisms of viral entry and budding within host cells. Here, we examined human airway epithelium infected with three different isolates of SARS-CoV-2 including the B.1.1.7 variant by transmission electron microscopy and tomography. For all isolates, the virus infected ciliated but not goblet epithelial cells. Key SARS-CoV-2 entry molecules, ACE2 and TMPRSS2, were found to be localised to the plasma membrane including microvilli but excluded from cilia. Consistently, extracellular virions were seen associated with microvilli and the apical plasma membrane but rarely with ciliary membranes. Profiles indicative of viral fusion where tomography showed that the viral membrane was continuous with the apical plasma membrane and the nucleocapsids diluted, compared with unfused virus, demonstrate that the plasma membrane is one site of entry where direct fusion releasing the nucleoprotein-encapsidated genome occurs. Intact intracellular virions were found within ciliated cells in compartments with a single membrane bearing S glycoprotein. Tomography showed concentration of nucleocapsids round the periphery of profiles strongly suggestive of viral budding into these compartments and this may explain how virions gain their S glycoprotein containing envelope.
History
DepositionFeb 16, 2022-
Header (metadata) releaseMar 23, 2022-
Map releaseMar 23, 2022-
UpdateApr 6, 2022-
Current statusApr 6, 2022Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

Downloads & links

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Map

FileDownload / File: emd_14367.map.gz / Format: CCP4 / Size: 687.7 MB / Type: IMAGE STORED AS SIGNED BYTE
AnnotationTomogram showing SARS-CoV-2 virions within a viral containing compartment of an infected airway cell
Voxel sizeX=Y=Z: 4.571 Å
Density
Minimum - Maximum-128.0 - 127.0
Average (Standard dev.)56.245956 (±15.635837)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin00-51
Dimensions26722672101
Spacing26722672101
CellA: 12213.712 Å / B: 12213.712 Å / C: 461.67102 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Airway epithelial cells infected with SARS-CoV-2

EntireName: Airway epithelial cells infected with SARS-CoV-2
Components
  • Cell: Airway epithelial cells infected with SARS-CoV-2

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Supramolecule #1: Airway epithelial cells infected with SARS-CoV-2

SupramoleculeName: Airway epithelial cells infected with SARS-CoV-2 / type: cell / ID: 1 / Parent: 0
Source (natural)Organism: Severe acute respiratory syndrome coronavirus 2

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Experimental details

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Structure determination

Methodnegative staining
Processingelectron tomography
Aggregation statecell

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Sample preparation

BufferpH: 7.4
StainingType: POSITIVE / Material: UA Zero
Sugar embeddingMaterial: Epon 812
SectioningUltramicrotomy - Instrument: Leica ultramicrotome / Ultramicrotomy - Temperature: 293 K / Ultramicrotomy - Final thickness: 100 nm

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Electron microscopy

MicroscopeJEOL 1400
Electron beamAcceleration voltage: 120 kV / Electron source: TUNGSTEN HAIRPIN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 0.7000000000000001 µm / Nominal defocus min: 0.7000000000000001 µm
Image recordingFilm or detector model: OTHER / Average electron dose: 50.0 e/Å2

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Image processing

Final reconstructionNumber images used: 120

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