Journal: Proc Natl Acad Sci U S A / Year: 2009 Title: Molecular interactions in rotavirus assembly and uncoating seen by high-resolution cryo-EM. Authors: James Z Chen / Ethan C Settembre / Scott T Aoki / Xing Zhang / A Richard Bellamy / Philip R Dormitzer / Stephen C Harrison / Nikolaus Grigorieff / Abstract: Rotaviruses, major causes of childhood gastroenteritis, are nonenveloped, icosahedral particles with double-strand RNA genomes. By the use of electron cryomicroscopy and single-particle ...Rotaviruses, major causes of childhood gastroenteritis, are nonenveloped, icosahedral particles with double-strand RNA genomes. By the use of electron cryomicroscopy and single-particle reconstruction, we have visualized a rotavirus particle comprising the inner capsid coated with the trimeric outer-layer protein, VP7, at a resolution (4 A) comparable with that of X-ray crystallography. We have traced the VP7 polypeptide chain, including parts not seen in its X-ray crystal structure. The 3 well-ordered, 30-residue, N-terminal "arms" of each VP7 trimer grip the underlying trimer of VP6, an inner-capsid protein. Structural differences between free and particle-bound VP7 and between free and VP7-coated inner capsids may regulate mRNA transcription and release. The Ca(2+)-stabilized VP7 intratrimer contact region, which presents important neutralizing epitopes, is unaltered upon capsid binding.
History
Deposition
Mar 25, 2009
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Header (metadata) release
Apr 8, 2009
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Map release
May 18, 2009
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Update
Mar 9, 2012
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Current status
Mar 9, 2012
Processing site: PDBe / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 90 K / Instrument: HOMEMADE PLUNGER / Details: Vitrification instrument: manual plunger / Method: Blot for 3 seconds before plunging
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Electron microscopy
Microscope
FEI TECNAI F30
Temperature
Min: 90 K / Max: 90 K / Average: 90 K
Date
Dec 1, 2007
Image recording
Category: FILM / Film or detector model: KODAK SO-163 FILM / Digitization - Scanner: ZEISS SCAI / Digitization - Sampling interval: 7 µm / Number real images: 148 / Average electron dose: 25 e/Å2 / Od range: 1.2 / Bits/pixel: 12
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
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