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- EMDB-44966: Sub-tomogram average of a pair of RSV F trimers from native virio... -

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Entry
Database: EMDB / ID: EMD-44966
TitleSub-tomogram average of a pair of RSV F trimers from native virions released from RSV-infected BEAS-2B cells cultured on EM grids
Map dataSub-tomogram average of a pair of RSV F trimers from native virions released from RSV-infected BEAS-2B cells cultured on EM grids
Sample
  • Virus: Human respiratory syncytial virus A2
KeywordsMembrane Fusion / VIRAL PROTEIN
Biological speciesHuman respiratory syncytial virus A2
Methodsubtomogram averaging / cryo EM / Resolution: 14.0 Å
AuthorsSibert BS / Wright ER
Funding support United States, 3 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R01 GM114561 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)U24 GM139168 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)U24 GM129547 United States
CitationJournal: Nat Commun / Year: 2024
Title: Assembly of respiratory syncytial virus matrix protein lattice and its coordination with fusion glycoprotein trimers.
Authors: Bryan S Sibert / Joseph Y Kim / Jie E Yang / Zunlong Ke / Christopher C Stobart / Martin L Moore / Elizabeth R Wright /
Abstract: Respiratory syncytial virus (RSV) is an enveloped, filamentous, negative-strand RNA virus that causes significant respiratory illness worldwide. RSV vaccines are available, however there is still ...Respiratory syncytial virus (RSV) is an enveloped, filamentous, negative-strand RNA virus that causes significant respiratory illness worldwide. RSV vaccines are available, however there is still significant need for research to support the development of vaccines and therapeutics against RSV and related Mononegavirales viruses. Individual virions vary in size, with an average diameter of ~130 nm and ranging from ~500 nm to over 10 µm in length. Though the general arrangement of structural proteins in virions is known, we use cryo-electron tomography and sub-tomogram averaging to determine the molecular organization of RSV structural proteins. We show that the peripheral membrane-associated RSV matrix (M) protein is arranged in a packed helical-like lattice of M-dimers. We report that RSV F glycoprotein is frequently observed as pairs of trimers oriented in an anti-parallel conformation to support potential interactions between trimers. Our sub-tomogram averages indicate the positioning of F-trimer pairs is correlated with the underlying M lattice. These results provide insight into RSV virion organization and may aid in the development of RSV vaccines and anti-viral targets.
History
DepositionMay 21, 2024-
Header (metadata) releaseJul 24, 2024-
Map releaseJul 24, 2024-
UpdateJul 24, 2024-
Current statusJul 24, 2024Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_44966.png

Downloads & links

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Map

FileDownload / File: emd_44966.map.gz / Format: CCP4 / Size: 144.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationSub-tomogram average of a pair of RSV F trimers from native virions released from RSV-infected BEAS-2B cells cultured on EM grids
Projections & slices

Image control

Size
Brightness
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Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.69 Å/pix.
x 336 pix.
= 568.848 Å		1.69 Å/pix.
x 336 pix.
= 568.848 Å		1.69 Å/pix.
x 336 pix.
= 568.848 Å

Surface

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Slices (1/2)

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Images are generated by Spider.

Voxel sizeX=Y=Z: 1.693 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.00765
Minimum - Maximum-0.010976217 - 0.020659396
Average (Standard dev.)0.00000000000003 (±0.0022747514)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions336336336
Spacing336336336
CellA=B=C: 568.84796 Å
α=β=γ: 90.0 °

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Supplemental data

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Mask #1

Fileemd_44966_msk_1.map
Projections & Slices
AxesZYX

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Half map: Half Map A

Fileemd_44966_half_map_1.map
AnnotationHalf Map A
Projections & Slices
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Histogram

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Half map: Half Map B

Fileemd_44966_half_map_2.map
AnnotationHalf Map B
Projections & Slices
AxesZYX

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Sample components

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Entire : Human respiratory syncytial virus A2

EntireName: Human respiratory syncytial virus A2
Components
  • Virus: Human respiratory syncytial virus A2

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Supramolecule #1: Human respiratory syncytial virus A2

SupramoleculeName: Human respiratory syncytial virus A2 / type: virus / ID: 1 / Parent: 0
Details: The strain was kindly provided by Dr. Martin L. Moore. The reference is PMCID: PMC3492879/PMID:23062737
NCBI-ID: 11259 / Sci species name: Human respiratory syncytial virus A2 / Sci species strain: rA2-mK+ / Virus type: VIRION / Virus isolate: STRAIN / Virus enveloped: Yes / Virus empty: No
Host (natural)Organism: Homo sapiens (human)

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Experimental details

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Structure determination

Methodcryo EM
Processingsubtomogram averaging
Aggregation stateparticle

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Sample preparation

BufferpH: 7.2 / Details: Cells were grown in RPMI medium.
GridModel: Quantifoil R2/1 / Material: GOLD / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY ARRAY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 60 sec.
VitrificationCryogen name: ETHANE / Chamber humidity: 80 % / Chamber temperature: 303 K / Instrument: GATAN CRYOPLUNGE 3
DetailsFilamentous virions released from BEAS-2B cells grown and infected with RSV A2-mk+ on cryo-TEM grids.

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Specialist opticsEnergy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Digitization - Dimensions - Width: 5760 pixel / Digitization - Dimensions - Height: 4096 pixel / Average exposure time: 0.35 sec. / Average electron dose: 2.44 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 6.0 µm / Nominal defocus min: 2.0 µm / Nominal magnification: 53000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 14.0 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 4.0) / Number subtomograms used: 21522
ExtractionNumber tomograms: 27 / Number images used: 305989
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION
FSC plot (resolution estimation)

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