|Entry||Database: PDB / ID: 6zts|
|Title||Assembly intermediates of orthoreovirus captured in the cell|
|Keywords||VIRUS LIKE PARTICLE / Assemble Intermediates / Orthoreovirus / cryo-electron tomography / cellular lamellae|
|Function / homology||Zinc finger C2H2-type / Lambda-1|
Function and homology information
|Biological species||Mammalian orthoreovirus|
|Method||ELECTRON MICROSCOPY / electron tomography / cryo EM|
|Authors||Sutton, G.C. / Stuart, D.I.|
|Funding support|| United Kingdom, 2items |
|Citation||Journal: Nat Commun / Year: 2020|
Title: Assembly intermediates of orthoreovirus captured in the cell.
Authors: Geoff Sutton / Dapeng Sun / Xiaofeng Fu / Abhay Kotecha / Corey W Hecksel / Daniel K Clare / Peijun Zhang / David I Stuart / Mark Boyce /
Abstract: Traditionally, molecular assembly pathways for viruses are inferred from high resolution structures of purified stable intermediates, low resolution images of cell sections and genetic approaches. ...Traditionally, molecular assembly pathways for viruses are inferred from high resolution structures of purified stable intermediates, low resolution images of cell sections and genetic approaches. Here, we directly visualise an unsuspected 'single shelled' intermediate for a mammalian orthoreovirus in cryo-preserved infected cells, by cryo-electron tomography of cellular lamellae. Particle classification and averaging yields structures to 5.6 Å resolution, sufficient to identify secondary structural elements and produce an atomic model of the intermediate, comprising 120 copies each of protein λ1 and σ2. This λ1 shell is 'collapsed' compared to the mature virions, with molecules pushed inwards at the icosahedral fivefolds by ~100 Å, reminiscent of the first assembly intermediate of certain prokaryotic dsRNA viruses. This supports the supposition that these viruses share a common ancestor, and suggests mechanisms for the assembly of viruses of the Reoviridae. Such methodology holds promise for dissecting the replication cycle of many viruses.
SummaryFull reportAbout validation report
|Structure viewer||Molecule: |
Downloads & links
Mass: 109603.016 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Mammalian orthoreovirus / References: UniProt: A0A023VYS9
|Experiment||Method: ELECTRON MICROSCOPY|
|EM experiment||Aggregation state: CELL / 3D reconstruction method: electron tomography|
|Component||Name: Assembly intermediates of orthoreovirus / Type: VIRUS / Entity ID: #1 / Source: NATURAL|
|Molecular weight||Experimental value: NO|
|Source (natural)||Organism: Mammalian orthoreovirus|
|Details of virus||Empty: YES / Enveloped: NO / Isolate: OTHER / Type: VIRION|
|Buffer solution||pH: 7|
|Specimen||Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES|
|Vitrification||Cryogen name: NITROGEN|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Microscopy||Model: FEI TITAN KRIOS|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM|
|Electron lens||Mode: DARK FIELD|
|Image recording||Electron dose: 2 e/Å2 / Film or detector model: GATAN K2 SUMMIT (4k x 4k)|
|CTF correction||Type: PHASE FLIPPING AND AMPLITUDE CORRECTION|
|3D reconstruction||Num. of particles: 41|
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- International Committee on Taxonomy of Viruses (ICTV) defined the short name of the 2019 coronavirus as "SARS-CoV-2".
The species Severe acute respiratory syndrome-related coronavirus: classifying 2019-nCoV and naming it SARS-CoV-2 - nature microbiology
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