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Assembly intermediates of orthoreovirus captured in the cell

DOI: 10.1038/s41467-020-18243-9 DOI Help

Authors: Geoff Sutton (Wellcome Centre for Human Genetics, University of Oxford) , Dapeng Sun (University of Pittsburgh) , Xiaofeng Fu (University of Pittsburgh) , Abhay Kotecha (Wellcome Centre for Human Genetics, University of Oxford) , Corey W. Hecksel (Diamond Light Source) , Daniel K. Clare (Diamond Light Source) , Peijun Zhang (Wellcome Centre for Human Genetics, University of Oxford; Diamond Light Source; University of Pittsburgh) , David I. Stuart (Wellcome Centre for Human Genetics, University of Oxford; Diamond Light Source) , Mark Boyce (Wellcome Centre for Human Genetics, University of Oxford)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Communications , VOL 11

State: Published (Approved)
Published: September 2020
Diamond Proposal Number(s): 18477

Open Access Open Access

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. 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.

Journal Keywords: Cryoelectron tomography; Virology

Diamond Keywords: Viruses

Subject Areas: Biology and Bio-materials

Diamond Offline Facilities: Electron Bio-Imaging Centre (eBIC)
Instruments: Krios III-Titan Krios III at Diamond


Discipline Tags:

Health & Wellbeing Infectious Diseases Life Sciences & Biotech Pathogens Structural biology

Technical Tags:

Cryo Electron Tomography (Cryo ET) Imaging Tomography