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- EMDB-17036: Double-headed nucleocapsid of the human Respiratory Syncytial Virus -

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Basic information

Entry
Database: EMDB / ID: EMD-17036
TitleDouble-headed nucleocapsid of the human Respiratory Syncytial Virus
Map dataDouble-headed nucleocapsid of the Respiratory Syncytial Virus.
Sample
  • Complex: Double-headed nucleocapsid of the human Respiratory Syncytial Virus
Keywordsspiral clam / N-RNA / nucleoprotein / nucleocapsid / VIRAL PROTEIN
Biological speciesHuman respiratory syncytial virus A strain Long
Methodsingle particle reconstruction / cryo EM / Resolution: 3.8 Å
AuthorsGonnin L / Desfosses A / Gutsche I
Funding support France, 1 items
OrganizationGrant numberCountry
Agence Nationale de la Recherche (ANR)ANR DecRisp ANR-19-CE11-0017-01 France
CitationJournal: Nat Commun / Year: 2023
Title: Structural landscape of the respiratory syncytial virus nucleocapsids.
Authors: Lorène Gonnin / Ambroise Desfosses / Maria Bacia-Verloop / Didier Chevret / Marie Galloux / Jean-François Éléouët / Irina Gutsche /
Abstract: Human Respiratory Syncytial Virus (HRSV) is a prevalent cause of severe respiratory infections in children and the elderly. The helical HRSV nucleocapsid is a template for the viral RNA synthesis and ...Human Respiratory Syncytial Virus (HRSV) is a prevalent cause of severe respiratory infections in children and the elderly. The helical HRSV nucleocapsid is a template for the viral RNA synthesis and a scaffold for the virion assembly. This cryo-electron microscopy analysis reveals the non-canonical arrangement of the HRSV nucleocapsid helix, composed of 16 nucleoproteins per asymmetric unit, and the resulting systematic variations in the RNA accessibility. We demonstrate that this unique helical symmetry originates from longitudinal interactions by the C-terminal arm of the HRSV nucleoprotein. We explore the polymorphism of the nucleocapsid-like assemblies, report five structures of the full-length particles and two alternative arrangements formed by a C-terminally truncated nucleoprotein mutant, and demonstrate the functional importance of the identified longitudinal interfaces. We put all these findings in the context of the HRSV RNA synthesis machinery and delineate the structural basis for its further investigation.
History
DepositionApr 6, 2023-
Header (metadata) releaseSep 27, 2023-
Map releaseSep 27, 2023-
UpdateSep 27, 2023-
Current statusSep 27, 2023Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

Downloads & links

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Map

FileDownload / File: emd_17036.map.gz / Format: CCP4 / Size: 325 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationDouble-headed nucleocapsid of the Respiratory Syncytial Virus.
Voxel sizeX=Y=Z: 1.145 Å
Density
Contour LevelBy AUTHOR: 0.11
Minimum - Maximum-0.5558655 - 1.0361762
Average (Standard dev.)0.0023359826 (±0.021564813)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions440440440
Spacing440440440
CellA=B=C: 503.8 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: Double-headed nucleocapsid of the Respiratory Syncytial Virus. Half...

Fileemd_17036_half_map_1.map
AnnotationDouble-headed nucleocapsid of the Respiratory Syncytial Virus. Half B.
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Half map: Double-headed nucleocapsid of the Respiratory Syncytial Virus. Half...

Fileemd_17036_half_map_2.map
AnnotationDouble-headed nucleocapsid of the Respiratory Syncytial Virus. Half A.
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Sample components

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Entire : Double-headed nucleocapsid of the human Respiratory Syncytial Virus

EntireName: Double-headed nucleocapsid of the human Respiratory Syncytial Virus
Components
  • Complex: Double-headed nucleocapsid of the human Respiratory Syncytial Virus

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Supramolecule #1: Double-headed nucleocapsid of the human Respiratory Syncytial Virus

SupramoleculeName: Double-headed nucleocapsid of the human Respiratory Syncytial Virus
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#2
Details: Double-headed nucleocapsid formed by the nucleoprotein N of the human RSV upon overexpression in insect cells and encapsidation of cellular RNA
Source (natural)Organism: Human respiratory syncytial virus A strain Long

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 7.4
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeTFS GLACIOS
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 2.4 µm / Nominal defocus min: 0.7000000000000001 µm
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 42.0 e/Å2

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Image processing

Startup modelType of model: NONE
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.8 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 25338

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