Journal: Cell / Year: 2010 Title: 3.3 A cryo-EM structure of a nonenveloped virus reveals a priming mechanism for cell entry. Authors: Xing Zhang / Lei Jin / Qin Fang / Wong H Hui / Z Hong Zhou / Abstract: To achieve cell entry, many nonenveloped viruses must transform from a dormant to a primed state. In contrast to the membrane fusion mechanism of enveloped viruses (e.g., influenza virus), this ...To achieve cell entry, many nonenveloped viruses must transform from a dormant to a primed state. In contrast to the membrane fusion mechanism of enveloped viruses (e.g., influenza virus), this membrane penetration mechanism is poorly understood. Here, using single-particle cryo-electron microscopy, we report a 3.3 A structure of the primed, infectious subvirion particle of aquareovirus. The density map reveals side-chain densities of all types of amino acids (except glycine), enabling construction of a full-atom model of the viral particle. Our structure and biochemical results show that priming involves autocleavage of the membrane penetration protein and suggest that Lys84 and Glu76 may facilitate this autocleavage in a nucleophilic attack. We observe a myristoyl group, covalently linked to the N terminus of the penetration protein and embedded in a hydrophobic pocket. These results suggest a well-orchestrated process of nonenveloped virus entry involving autocleavage of the penetration protein prior to exposure of its membrane-insertion finger.
History
Deposition
Jan 18, 2010
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Header (metadata) release
May 3, 2010
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Map release
May 3, 2010
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Update
Dec 26, 2012
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Current status
Dec 26, 2012
Processing site: RCSB / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
Name: Aquareovirus / type: sample / ID: 1000 / Details: The sample was monodisperse / Number unique components: 4
Molecular weight
Experimental: 72 MDa / Theoretical: 72 MDa
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Supramolecule #1: Aquareovirus
Supramolecule
Name: Aquareovirus / type: virus / ID: 1 / Name.synonym: GCRV / NCBI-ID: 10979 / Sci species name: Aquareovirus / Database: NCBI / Virus type: OTHER / Virus isolate: STRAIN / Virus enveloped: No / Virus empty: No / Syn species name: GCRV
Host (natural)
synonym: VERTEBRATES
Molecular weight
Experimental: 72 MDa / Theoretical: 72 MDa
Virus shell
Shell ID: 1 / Name: ISVP / Diameter: 800 Å / T number (triangulation number): 13
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Experimental details
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Structure determination
Method
cryo EM
Processing
single particle reconstruction
Aggregation state
particle
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Sample preparation
Buffer
pH: 7.5 / Details: 10mM PBS Buffer
Grid
Details: 400 mesh quantifoil 1.2/1.3
Vitrification
Cryogen name: METHANE / Chamber humidity: 100 % / Chamber temperature: 90 K / Instrument: OTHER / Details: Vitrification instrument: FEI Vitrobot / Method: Blot for 7-9 seconds before plunging
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Electron microscopy
Microscope
FEI TITAN KRIOS
Temperature
Min: 90 K / Average: 90 K
Alignment procedure
Legacy - Astigmatism: objective lens astigmatism was corrected at 250,000 times magnification
Date
Mar 1, 2009
Image recording
Category: CCD / Film or detector model: KODAK SO-163 FILM / Digitization - Sampling interval: 6.35 µm / Number real images: 700 / Average electron dose: 25 e/Å2 / Bits/pixel: 16
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
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