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- EMDB-10097: Faba bean necrotic stunt virus (FBNSV) -

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Basic information

Entry
Database: EMDB / ID: EMD-10097
TitleFaba bean necrotic stunt virus (FBNSV)
Map datasharpened map
Sample
  • Virus: Faba bean necrotic stunt virus
    • Protein or peptide: Capsid proteinCapsid
Function / homologyNanovirus coat protein / Nanovirus coat protein / virion component => GO:0044423 / Capsid protein
Function and homology information
Biological speciesFaba bean necrotic stunt virus
Methodsingle particle reconstruction / cryo EM / Resolution: 3.19 Å
AuthorsTrapani S / Lai Kee Him J / Blanc S / Bron P
Funding support France, 1 items
OrganizationGrant numberCountry
French National Research AgencyANR-14- 15 CE02-0014 France
CitationJournal: PLoS Pathog / Year: 2023
Title: Structure-guided mutagenesis of the capsid protein indicates that a nanovirus requires assembled viral particles for systemic infection.
Authors: Stefano Trapani / Eijaz Ahmed Bhat / Michel Yvon / Joséphine Lai-Kee-Him / François Hoh / Marie-Stéphanie Vernerey / Elodie Pirolles / Mélia Bonnamy / Guy Schoehn / Jean-Louis Zeddam / ...Authors: Stefano Trapani / Eijaz Ahmed Bhat / Michel Yvon / Joséphine Lai-Kee-Him / François Hoh / Marie-Stéphanie Vernerey / Elodie Pirolles / Mélia Bonnamy / Guy Schoehn / Jean-Louis Zeddam / Stéphane Blanc / Patrick Bron /
Abstract: Nanoviruses are plant multipartite viruses with a genome composed of six to eight circular single-stranded DNA segments. The distinct genome segments are encapsidated individually in icosahedral ...Nanoviruses are plant multipartite viruses with a genome composed of six to eight circular single-stranded DNA segments. The distinct genome segments are encapsidated individually in icosahedral particles that measure ≈18 nm in diameter. Recent studies on the model species Faba bean necrotic stunt virus (FBNSV) revealed that complete sets of genomic segments rarely occur in infected plant cells and that the function encoded by a given viral segment can complement the others across neighbouring cells, presumably by translocation of the gene products through unknown molecular processes. This allows the viral genome to replicate, assemble into viral particles and infect anew, even with the distinct genome segments scattered in different cells. Here, we question the form under which the FBNSV genetic material propagates long distance within the vasculature of host plants and, in particular, whether viral particle assembly is required. Using structure-guided mutagenesis based on a 3.2 Å resolution cryogenic-electron-microscopy reconstruction of the FBNSV particles, we demonstrate that specific site-directed mutations preventing capsid formation systematically suppress FBNSV long-distance movement, and thus systemic infection of host plants, despite positive detection of the mutated coat protein when the corresponding segment is agroinfiltrated into plant leaves. These results strongly suggest that the viral genome does not propagate within the plant vascular system under the form of uncoated DNA molecules or DNA:coat-protein complexes, but rather moves long distance as assembled viral particles.
History
DepositionJun 26, 2019-
Header (metadata) releaseJul 17, 2019-
Map releaseJul 15, 2020-
UpdateFeb 1, 2023-
Current statusFeb 1, 2023Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.0162
  • Imaged by UCSF Chimera
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  • Surface view colored by radius
  • Surface level: 0.0162
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-6s44
  • Surface level: 0.0162
  • Imaged by UCSF Chimera
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  • Simplified surface model + fitted atomic model
  • Atomic modelsPDB-6s44
  • Imaged by Jmol
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

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Map

FileDownload / File: emd_10097.map.gz / Format: CCP4 / Size: 172.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotationsharpened map
Voxel sizeX=Y=Z: 1.21 Å
Density
Contour LevelBy AUTHOR: 0.0162 / Movie #1: 0.0162
Minimum - Maximum-0.03368195 - 0.07579645
Average (Standard dev.)5.2229225e-05 (±0.0035383527)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions356356356
Spacing356356356
CellA=B=C: 430.76 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.211.211.21
M x/y/z356356356
origin x/y/z0.0000.0000.000
length x/y/z430.760430.760430.760
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS356356356
D min/max/mean-0.0340.0760.000

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Supplemental data

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Mask #1

Fileemd_10097_msk_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Additional map: unsharpened map

Fileemd_10097_additional.map
Annotationunsharpened map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Half map: half dataset map 2

Fileemd_10097_half_map_1.map
Annotationhalf dataset map 2
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Half map: half dataset map 1

Fileemd_10097_half_map_2.map
Annotationhalf dataset map 1
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Sample components

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Entire : Faba bean necrotic stunt virus

EntireName: Faba bean necrotic stunt virus
Components
  • Virus: Faba bean necrotic stunt virus
    • Protein or peptide: Capsid proteinCapsid

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Supramolecule #1: Faba bean necrotic stunt virus

SupramoleculeName: Faba bean necrotic stunt virus / type: virus / ID: 1 / Parent: 0 / Macromolecule list: all / NCBI-ID: 283824 / Sci species name: Faba bean necrotic stunt virus / Virus type: VIRION / Virus isolate: STRAIN / Virus enveloped: No / Virus empty: No
Virus shellShell ID: 1 / Name: capsid / T number (triangulation number): 3

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Macromolecule #1: Capsid protein

MacromoleculeName: Capsid protein / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Faba bean necrotic stunt virus
Molecular weightTheoretical: 19.200422 KDa
SequenceString:
MVSNWNWSGK KGRRTPRRGY TRPFKSAVPT TRVVVHQSAV LKKDDVSGSE IKPEGDVARY KIRKVMLSCT LRMRPGELVN YLIVKCSSP IVNWSAAFTA PALMVKESCQ DMITIIGKGK VESNGVAGSD CTKSFNKFIR LGAGISQTQH LYVVMYTSEA V KTVLEHRV YIEV

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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
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI POLARA 300
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 40.0 e/Å2
Experimental equipment
Model: Tecnai Polara / Image courtesy: FEI Company

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Image processing

Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3)
Final reconstructionNumber classes used: 1 / Applied symmetry - Point group: I (icosahedral) / Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 3.19 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3) / Number images used: 5156
FSC plot (resolution estimation)

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Atomic model buiding 1

RefinementSpace: REAL / Protocol: AB INITIO MODEL
Output model

PDB-6s44:
Faba bean necrotic stunt virus (FBNSV)

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