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- EMDB-3284: Cryo-EM structure of BK polyomavirus VP1 virus-like particle -

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Basic information

Entry
Database: EMDB / ID: EMD-3284
TitleCryo-EM structure of BK polyomavirus VP1 virus-like particle
Map dataReconstruction of BK VP1 VLP (sharpened/masked)
Sample
  • Sample: BK polyomavirus VP1 VLP
  • Virus: BK polyomavirus VP1 VLP
KeywordsBKPyV / BK / polyomavirus / virus-like particle / VLP
Biological speciesBK polyomavirus VP1 VLP
Methodsingle particle reconstruction / cryo EM / Resolution: 9.12 Å
AuthorsHurdiss DL / Morgan EL / Thompson RF / Prescott EL / Panou MM / Macdonald A / Ranson NA
CitationJournal: Structure / Year: 2016
Title: New Structural Insights into the Genome and Minor Capsid Proteins of BK Polyomavirus using Cryo-Electron Microscopy.
Authors: Daniel L Hurdiss / Ethan L Morgan / Rebecca F Thompson / Emma L Prescott / Margarita M Panou / Andrew Macdonald / Neil A Ranson /
Abstract: BK polyomavirus is the causative agent of several diseases in transplant patients and the immunosuppressed. In order to better understand the structure and life cycle of BK, we produced infectious ...BK polyomavirus is the causative agent of several diseases in transplant patients and the immunosuppressed. In order to better understand the structure and life cycle of BK, we produced infectious virions and VP1-only virus-like particles in cell culture, and determined their three-dimensional structures using cryo-electron microscopy (EM) and single-particle image processing. The resulting 7.6-Å resolution structure of BK and 9.1-Å resolution of the virus-like particles are the highest-resolution cryo-EM structures of any polyomavirus. These structures confirm that the architecture of the major structural protein components of these human polyomaviruses are similar to previous structures from other hosts, but give new insight into the location and role of the enigmatic minor structural proteins, VP2 and VP3. We also observe two shells of electron density, which we attribute to a structurally ordered part of the viral genome, and discrete contacts between this density and both VP1 and the minor capsid proteins.
History
SupersessionID: EMD-3002
DepositionDec 17, 2015-
Header (metadata) releaseJan 13, 2016-
Map releaseApr 27, 2016-
UpdateMay 18, 2016-
Current statusMay 18, 2016Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.009
  • Imaged by UCSF Chimera
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  • Surface view colored by radius
  • Surface level: 0.009
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_3284.tif

Downloads & links

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Map

FileDownload / File: emd_3284.map.gz / Format: CCP4 / Size: 465.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationReconstruction of BK VP1 VLP (sharpened/masked)
Voxel sizeX=Y=Z: 1.35 Å
Density
Contour LevelBy AUTHOR: 0.009 / Movie #1: 0.009
Minimum - Maximum-0.00716442 - 0.01983405
Average (Standard dev.)0.00076196 (±0.00246748)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin-249-249-249
Dimensions500500500
Spacing500500500
CellA=B=C: 675.0 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.351.351.35
M x/y/z500500500
origin x/y/z0.0000.0000.000
length x/y/z675.000675.000675.000
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS-249-249-249
NC/NR/NS500500500
D min/max/mean-0.0070.0200.001

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Supplemental data

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Supplemental map: EMD-3284 unsharpened.map

FileEMD-3284_unsharpened.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 : BK polyomavirus VP1 VLP

EntireName: BK polyomavirus VP1 VLP
Components
  • Sample: BK polyomavirus VP1 VLP
  • Virus: BK polyomavirus VP1 VLP

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Supramolecule #1000: BK polyomavirus VP1 VLP

SupramoleculeName: BK polyomavirus VP1 VLP / type: sample / ID: 1000 / Oligomeric state: Icosohedral / Number unique components: 1

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Supramolecule #1: BK polyomavirus VP1 VLP

SupramoleculeName: BK polyomavirus VP1 VLP / type: virus / ID: 1 / Sci species name: BK polyomavirus VP1 VLP / Sci species strain: BKV-Ia / Virus type: VIRUS-LIKE PARTICLE / Virus isolate: STRAIN / Virus enveloped: No / Virus empty: No
Host (natural)Organism: Homo sapiens (human) / synonym: VERTEBRATES
Host systemRecombinant cell: HEK293TT / Recombinant plasmid: pIaw
Virus shellShell ID: 1 / Diameter: 498 Å

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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: 7.9 / Details: 10mM HEPES pH 7.9, 50mM CaCl2, 1mM MgCl2, 5mM KCl
GridDetails: Quantifoil R2/1 EM grids
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK IV / Method: 6.5 seconds blot before plunging

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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 / Nominal defocus max: 6.136 µm / Nominal defocus min: 0.526 µm / Nominal magnification: 19000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER
DateMay 13, 2015
Image recordingCategory: CCD / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Number real images: 170 / Average electron dose: 40 e/Å2
Details: 4 e-/A2/s, a 4 frames per second frame rate, and a 10 s exposure
Tilt angle min0
Tilt angle max0
Experimental equipment
Model: Tecnai Polara / Image courtesy: FEI Company

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Image processing

CTF correctionDetails: CTFFIND3
Final reconstructionApplied symmetry - Point group: I (icosahedral) / Resolution.type: BY AUTHOR / Resolution: 9.12 Å / Resolution method: OTHER / Software - Name: Relion / Number images used: 2888
FSC plot (resolution estimation)

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