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- EMDB-0050: DNA-devoid HCIV-1 virus particle -

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Basic information

Entry
Database: EMDB / ID: EMD-0050
TitleDNA-devoid HCIV-1 virus particle
Map dataThis map corresponds to DNA-devoid HCIV-1 particles. A cut through shows the lack of DNA and the unexpanded membrane vesicle. To better distinguish the membrane it is advisable to filter to 10 Ang.
Sample
  • Virus: Haloarcula californiae ATCC 33799 (Halophile)
Biological speciesHaloarcula californiae ATCC 33799 (Halophile)
Methodsingle particle reconstruction / cryo EM / Resolution: 14.9 Å
AuthorsSantos-Perez I / Charro D / Gil-Carton D / Azkargorta M / Elortza F / Bamford DH / Oksanen HM / Abrescia NGA
CitationJournal: Nat Commun / Year: 2019
Title: Structural basis for assembly of vertical single β-barrel viruses.
Authors: Isaac Santos-Pérez / Diego Charro / David Gil-Carton / Mikel Azkargorta / Felix Elortza / Dennis H Bamford / Hanna M Oksanen / Nicola G A Abrescia /
Abstract: The vertical double β-barrel major capsid protein (MCP) fold, fingerprint of the PRD1-adeno viral lineage, is widespread in many viruses infecting organisms across the three domains of life. The ...The vertical double β-barrel major capsid protein (MCP) fold, fingerprint of the PRD1-adeno viral lineage, is widespread in many viruses infecting organisms across the three domains of life. The discovery of PRD1-like viruses with two MCPs challenged the known assembly principles. Here, we present the cryo-electron microscopy (cryo-EM) structures of the archaeal, halophilic, internal membrane-containing Haloarcula californiae icosahedral virus 1 (HCIV-1) and Haloarcula hispanica icosahedral virus 2 (HHIV-2) at 3.7 and 3.8 Å resolution, respectively. Our structures reveal proteins located beneath the morphologically distinct two- and three-tower capsomers and homopentameric membrane proteins at the vertices that orchestrate the positioning of pre-formed vertical single β-barrel MCP heterodimers. The cryo-EM based structures together with the proteomics data provide insights into the assembly mechanism of this type of viruses and into those with membrane-less double β-barrel MCPs.
History
DepositionJun 13, 2018-
Header (metadata) releaseAug 22, 2018-
Map releaseMar 27, 2019-
UpdateApr 3, 2019-
Current statusApr 3, 2019Processing site: PDBe / Status: Released

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Structure visualization

Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_0050.png

Downloads & links

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Map

FileDownload / File: emd_0050.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationThis map corresponds to DNA-devoid HCIV-1 particles. A cut through shows the lack of DNA and the unexpanded membrane vesicle. To better distinguish the membrane it is advisable to filter to 10 Ang.
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
2.8 Å/pix.
x 384 pix.
= 1075.2 Å		2.8 Å/pix.
x 384 pix.
= 1075.2 Å		2.8 Å/pix.
x 384 pix.
= 1075.2 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 2.8 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.135
Minimum - Maximum-0.31705016 - 1.104037
Average (Standard dev.)0.0006336519 (±0.042282388)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions384384384
Spacing384384384
CellA=B=C: 1075.2 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #2

Fileemd_0050_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #1

Fileemd_0050_half_map_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 : Haloarcula californiae ATCC 33799

EntireName: Haloarcula californiae ATCC 33799 (Halophile)
Components
  • Virus: Haloarcula californiae ATCC 33799 (Halophile)

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Supramolecule #1: Haloarcula californiae ATCC 33799

SupramoleculeName: Haloarcula californiae ATCC 33799 / type: virus / ID: 1 / Parent: 0 / Details: Haloarcula californiae icosahedral virus - 1 / NCBI-ID: 662475 / Sci species name: Haloarcula californiae ATCC 33799 / Virus type: VIRION / Virus isolate: SPECIES / Virus enveloped: No / Virus empty: Yes
Host (natural)Organism: Haloarcula californiae ATCC 33799 (Halophile)
Virus shellShell ID: 1 / Diameter: 800.0 Å / T number (triangulation number): 28

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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

Concentration1.2 mg/mL
BufferpH: 7.2
Component:
ConcentrationFormulaName
1.0 MNaClSodium Chloride
70.0 mMMgCl2Magnesium Chloride
20.0 mMKClPotasium Chloride
1.0 mMCaCl2Calcium Chloride
50.0 mMC4H11NO3Tris-HCl
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: PLASMA CLEANING
VitrificationCryogen name: ETHANE / Instrument: FEI VITROBOT MARK III

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: FEI FALCON II (4k x 4k) / Detector mode: INTEGRATING / Digitization - Frames/image: 1-26 / Number grids imaged: 1 / Number real images: 3218 / Average electron dose: 36.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 3.9 µm / Nominal defocus min: 0.6 µm
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

Particle selectionNumber selected: 4584
CTF correctionSoftware - Name: CTFFIND (ver. 4)
Startup modelType of model: OTHER / Details: Associate ENTRY (HCIV-1).
Final reconstructionNumber classes used: 1 / Applied symmetry - Point group: I (icosahedral) / Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 14.9 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: RELION (ver. 2.0) / Number images used: 84
Initial angle assignmentType: PROJECTION MATCHING / Software - Name: RELION (ver. 2.0)
Final angle assignmentType: PROJECTION MATCHING / Software - Name: RELION (ver. 2.0)
Final 3D classificationNumber classes: 10 / Software - Name: RELION (ver. 2.0)
FSC plot (resolution estimation)

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