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- EMDB-15345: CryoEM structure of sweet potato feathery mottle virus VLP -

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Basic information

Entry
Database: EMDB / ID: EMD-15345
TitleCryoEM structure of sweet potato feathery mottle virus VLP
Map dataSharpened map.
Sample
  • Virus: Sweet potato feathery mottle virus
    • Protein or peptide: Genome polyprotein
    • RNA: Single-stranded RNARNA
Function / homologyPotyvirus coat protein / Potyvirus coat protein / viral capsid / Genome polyprotein
Function and homology information
Biological speciesSweet potato feathery mottle virus / Ipomoea batatas (sweet potato)
Methodhelical reconstruction / cryo EM / Resolution: 2.6 Å
AuthorsJaved A / Byrne JM / Ranson N / Lomonosoff G
Funding support Spain, United Kingdom, 4 items
OrganizationGrant numberCountry
Ministerio de Ciencia e Innovacion (MCIN)PID2019-105692RB-100 Spain
Ministerio de Ciencia e Innovacion (MCIN)CEX2019-000902-S Spain
Biotechnology and Biological Sciences Research Council (BBSRC)BB/R001669/1 United Kingdom
Biotechnology and Biological Sciences Research Council (BBSRC)BB/T004703/1 United Kingdom
CitationJournal: Commun Biol / Year: 2023
Title: CryoEM and stability analysis of virus-like particles of potyvirus and ipomovirus infecting a common host.
Authors: Ornela Chase / Abid Javed / Matthew J Byrne / Eva C Thuenemann / George P Lomonossoff / Neil A Ranson / Juan José López-Moya /
Abstract: Sweet potato feathery mottle virus (SPFMV) and Sweet potato mild mottle virus (SPMMV) are members of the genera Potyvirus and Ipomovirus, family Potyviridae, sharing Ipomoea batatas as common host, ...Sweet potato feathery mottle virus (SPFMV) and Sweet potato mild mottle virus (SPMMV) are members of the genera Potyvirus and Ipomovirus, family Potyviridae, sharing Ipomoea batatas as common host, but transmitted, respectively, by aphids and whiteflies. Virions of family members consist of flexuous rods with multiple copies of a single coat protein (CP) surrounding the RNA genome. Here we report the generation of virus-like particles (VLPs) by transient expression of the CPs of SPFMV and SPMMV in the presence of a replicating RNA in Nicotiana benthamiana. Analysis of the purified VLPs by cryo-electron microscopy, gave structures with resolutions of 2.6 and 3.0 Å, respectively, showing a similar left-handed helical arrangement of 8.8 CP subunits per turn with the C-terminus at the inner surface and a binding pocket for the encapsidated ssRNA. Despite their similar architecture, thermal stability studies reveal that SPMMV VLPs are more stable than those of SPFMV.
History
DepositionJul 5, 2022-
Header (metadata) releaseMay 17, 2023-
Map releaseMay 17, 2023-
UpdateMay 17, 2023-
Current statusMay 17, 2023Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_15345.png

Downloads & links

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Map

FileDownload / File: emd_15345.map.gz / Format: CCP4 / Size: 34.3 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationSharpened map.
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.07 Å/pix.
x 208 pix.
= 221.52 Å		1.07 Å/pix.
x 208 pix.
= 221.52 Å		1.07 Å/pix.
x 208 pix.
= 221.52 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.065 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.03
Minimum - Maximum-0.16347069 - 0.31914026
Average (Standard dev.)0.0015139481 (±0.018838866)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions208208208
Spacing208208208
CellA=B=C: 221.52002 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: Half 1 map.

Fileemd_15345_half_map_1.map
AnnotationHalf 1 map.
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: Half 2 map

Fileemd_15345_half_map_2.map
AnnotationHalf 2 map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Sweet potato feathery mottle virus

EntireName: Sweet potato feathery mottle virus
Components
  • Virus: Sweet potato feathery mottle virus
    • Protein or peptide: Genome polyprotein
    • RNA: Single-stranded RNARNA

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Supramolecule #1: Sweet potato feathery mottle virus

SupramoleculeName: Sweet potato feathery mottle virus / type: virus / ID: 1 / Parent: 0 / Macromolecule list: all / NCBI-ID: 12844 / Sci species name: Sweet potato feathery mottle virus / Virus type: VIRUS-LIKE PARTICLE / Virus isolate: SPECIES / Virus enveloped: No / Virus empty: No
Host (natural)Organism: Ipomoea batatas (sweet potato)
Molecular weightTheoretical: 380 KDa
Virus shellShell ID: 1 / Diameter: 130.0 Å

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Macromolecule #1: Genome polyprotein

MacromoleculeName: Genome polyprotein / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Sweet potato feathery mottle virus
Molecular weightTheoretical: 25.209426 KDa
Recombinant expressionOrganism: Nicotiana benthamiana (plant)
SequenceString: VPRVKMNANK KRQPMVNGRA IINFQHLSTY EPEQFEVANT RSTQEQFQAW YEGVKGDYGV DDTGMGILLN GLMVWCIENG TSPNINGVW TMMDGDEQVT YPIKPLLDHA VPTFRQIMTH FSDVAEAYIE MRNRTKAYMP RYGLQRNLTD MSLARYAFDF Y ELHSTTPA ...String:
VPRVKMNANK KRQPMVNGRA IINFQHLSTY EPEQFEVANT RSTQEQFQAW YEGVKGDYGV DDTGMGILLN GLMVWCIENG TSPNINGVW TMMDGDEQVT YPIKPLLDHA VPTFRQIMTH FSDVAEAYIE MRNRTKAYMP RYGLQRNLTD MSLARYAFDF Y ELHSTTPA RAKEAHLQMK AAALKNARNR LFGLDGNVST QEEDTERHTT TDVTRNIHNL LG

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Macromolecule #2: Single-stranded RNA

MacromoleculeName: Single-stranded RNA / type: rna / ID: 2 / Number of copies: 1
Source (natural)Organism: Ipomoea batatas (sweet potato)
Molecular weightTheoretical: 1.485872 KDa
SequenceString:
UUUUU

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Experimental details

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Structure determination

Methodcryo EM
Processinghelical reconstruction
Aggregation statefilament

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Sample preparation

BufferpH: 7 / Component - Concentration: 50.0 mM / Component - Formula: NaPO4 / Component - Name: Sodium Phosphate
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec.
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: 3.0 µm / Nominal defocus min: 0.8 µm
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Image recordingFilm or detector model: FEI FALCON III (4k x 4k) / Detector mode: COUNTING / Number grids imaged: 1 / Number real images: 3276 / Average electron dose: 1.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Segment selectionNumber selected: 313330
Startup modelType of model: OTHER / Details: Cylinder with a diameter of the filament.
Final angle assignmentType: NOT APPLICABLE / Software - Name: RELION (ver. 3.1)
Final reconstructionNumber classes used: 1
Applied symmetry - Helical parameters - Δz: 3.98 Å
Applied symmetry - Helical parameters - Δ&Phi: 40.93 °
Applied symmetry - Helical parameters - Axial symmetry: C1 (asymmetric)
Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 2.6 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.1) / Number images used: 48042
FSC plot (resolution estimation)

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Atomic model buiding 1

RefinementSpace: REAL / Protocol: FLEXIBLE FIT / Overall B value: 140.9 / Target criteria: Correlation coefficient
Output model

PDB-8acb:
CryoEM structure of sweet potato feathery mottle virus VLP

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