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- EMDB-20430: Kaposi's sarcoma-associated herpesvirus (KSHV), C1 virion capsid ... -

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Basic information

Entry
Database: EMDB / ID: EMD-20430
TitleKaposi's sarcoma-associated herpesvirus (KSHV), C1 virion capsid reconstruction
Map data
SampleHuman gammaherpesvirus 8:
virus
Biological speciesHuman gammaherpesvirus 8
Methodsingle particle reconstruction / cryo EM / Resolution: 7.6 Å
AuthorsGong D / Dai X / Jih J / Liu YT / Bi GQ / Sun R / Zhou ZH
Funding support United States, China, 14 items
OrganizationGrant numberCountry
National Institutes of Health/National Center for Research Resources1S10RR23057 United States
Other government2016YFA0400900 China
National Science Foundation (United States)DBI-1338135 United States
National Institutes of Health/National Institute of General Medical Sciences1U24GM116792 United States
National Science Foundation (United States)DMR-1548924 United States
National Institutes of Health/National Institute of General Medical SciencesGM071940 United States
National Institutes of Health/National Institute of Dental and Craniofacial ResearchDE028583 United States
National Institutes of Health/National Institute of Dental and Craniofacial ResearchDE025567 United States
National Institutes of Health/National Institute Of Allergy and Infectious DiseasesAI094386 United States
National Institutes of Health/National Institute of Dental and Craniofacial ResearchDE027901 United States
National Institutes of Health/National Cancer InstituteCA091791 United States
National Institutes of Health/National Cancer InstituteCA177322 United States
Other government2017YFA0505300 China
National Institutes of Health/National Institute of Dental and Craniofacial ResearchDE023591 United States
CitationJournal: Cell / Year: 2019
Title: DNA-Packing Portal and Capsid-Associated Tegument Complexes in the Tumor Herpesvirus KSHV.
Authors: Danyang Gong / Xinghong Dai / Jonathan Jih / Yun-Tao Liu / Guo-Qiang Bi / Ren Sun / Z Hong Zhou /
Abstract: Assembly of Kaposi's sarcoma-associated herpesvirus (KSHV) begins at a bacteriophage-like portal complex that nucleates formation of an icosahedral capsid with capsid-associated tegument complexes ...Assembly of Kaposi's sarcoma-associated herpesvirus (KSHV) begins at a bacteriophage-like portal complex that nucleates formation of an icosahedral capsid with capsid-associated tegument complexes (CATCs) and facilitates translocation of an ∼150-kb dsDNA genome, followed by acquisition of a pleomorphic tegument and envelope. Because of deviation from icosahedral symmetry, KSHV portal and tegument structures have largely been obscured in previous studies. Using symmetry-relaxed cryo-EM, we determined the in situ structure of the KSHV portal and its interactions with surrounding capsid proteins, CATCs, and the terminal end of KSHV's dsDNA genome. Our atomic models of the portal and capsid/CATC, together with visualization of CATCs' variable occupancy and alternate orientation of CATC-interacting vertex triplexes, suggest a mechanism whereby the portal orchestrates procapsid formation and asymmetric long-range determination of CATC attachment during DNA packaging prior to pleomorphic tegumentation/envelopment. Structure-based mutageneses confirm that a triplex deep binding groove for CATCs is a hotspot that holds promise for antiviral development.
History
DepositionJul 6, 2019-
Header (metadata) releaseJul 31, 2019-
Map releaseSep 11, 2019-
UpdateSep 18, 2019-
Current statusSep 18, 2019Processing site: RCSB / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.014
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by radius
  • Surface level: 0.014
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

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Map

FileDownload / File: emd_20430.map.gz / Format: CCP4 / Size: 1.4 GB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
2.06 Å/pix.
x 720 pix.
= 1483.2 Å
2.06 Å/pix.
x 720 pix.
= 1483.2 Å
2.06 Å/pix.
x 720 pix.
= 1483.2 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 2.06 Å
Density
Contour LevelBy AUTHOR: 0.014 / Movie #1: 0.014
Minimum - Maximum-0.024108846 - 0.029338676
Average (Standard dev.)0.0007912924 (±0.0032670132)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions720720720
Spacing720720720
CellA=B=C: 1483.2 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z2.062.062.06
M x/y/z720720720
origin x/y/z0.0000.0000.000
length x/y/z1483.2001483.2001483.200
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS720720720
D min/max/mean-0.0180.0350.002

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Supplemental data

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Sample components

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Entire Human gammaherpesvirus 8

EntireName: Human gammaherpesvirus 8 / Number of components: 1

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Component #1: virus, Human gammaherpesvirus 8

VirusName: Human gammaherpesvirus 8Kaposi's sarcoma-associated herpesvirus
Class: VIRION / Empty: No / Enveloped: Yes / Isolate: STRAIN
SpeciesSpecies: Human gammaherpesvirus 8 / Strain: BAC16
Source (natural)Host Species: Homo sapiens (human)
Shell #1Name of element: Capsid / Diameter: 1250.0 Å / T number (triangulation number): 16

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Experimental details

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Sample preparation

SpecimenSpecimen state: Particle / Method: cryo EM
Sample solutionpH: 7.4
VitrificationInstrument: HOMEMADE PLUNGER / Cryogen name: ETHANE / Temperature: 298 K
Details: The sample was manually blotted and frozen with a homemade plunger..

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
ImagingMicroscope: FEI TITAN KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 25 e/Å2 / Illumination mode: FLOOD BEAM
LensMagnification: 14000.0 X (nominal), 24271.0 X (calibrated) / Cs: 2.7 mm / Imaging mode: BRIGHT FIELD
Specimen HolderModel: FEI TITAN KRIOS AUTOGRID HOLDER / Temperature: ( - 79.0 K)
CameraDetector: GATAN K2 SUMMIT (4k x 4k)

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Image acquisition

Image acquisitionNumber of digital images: 8007 / Sampling size: 2.5 µm

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Image processing

ProcessingMethod: single particle reconstruction / Applied symmetry: C1 (asymmetric) / Number of projections: 39073
3D reconstructionAlgorithm: FOURIER SPACE / Software: RELION / Resolution: 7.6 Å / Resolution method: FSC 0.143 CUT-OFF
FSC plot (resolution estimation)

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