Cryo-electron Microscopy Structure, Assembly, and Mechanics Show Morphogenesis and Evolution of Human Picobirnavirus.

Autor:	Ortega-Esteban Á; Department of Structure of Macromolecules, Centro Nacional de Biotecnología, Madrid, Spain., Mata CP; Department of Structure of Macromolecules, Centro Nacional de Biotecnología, Madrid, Spain., Rodríguez-Espinosa MJ; Department of Structure of Macromolecules, Centro Nacional de Biotecnología, Madrid, Spain., Luque D; Centro Nacional de Microbiología/ISCIII, Majadahonda, Madrid, Spain., Irigoyen N; Department of Structure of Macromolecules, Centro Nacional de Biotecnología, Madrid, Spain., Rodríguez JM; Department of Structure of Macromolecules, Centro Nacional de Biotecnología, Madrid, Spain., de Pablo PJ; Department of Condensed Matter Physics, Universidad Autónoma de Madrid, Madrid, Spain., Castón JR; Department of Structure of Macromolecules, Centro Nacional de Biotecnología, Madrid, Spain jrcaston@cnb.csic.es.
Jazyk:	angličtina
Zdroj:	Journal of virology [J Virol] 2020 Nov 23; Vol. 94 (24). Date of Electronic Publication: 2020 Nov 23 (Print Publication: 2020).
DOI:	10.1128/JVI.01542-20
Abstrakt:	Despite their diversity, most double-stranded-RNA (dsRNA) viruses share a specialized T=1 capsid built from dimers of a single protein that provides a platform for genome transcription and replication. This ubiquitous capsid remains structurally undisturbed throughout the viral cycle, isolating the genome to avoid triggering host defense mechanisms. Human picobirnavirus (hPBV) is a dsRNA virus frequently associated with gastroenteritis, although its pathogenicity is yet undefined. Here, we report the cryo-electron microscopy (cryo-EM) structure of hPBV at 2.6-Å resolution. The capsid protein (CP) is arranged in a single-shelled, ∼380-Å-diameter T=1 capsid with a rough outer surface similar to that of dsRNA mycoviruses. The hPBV capsid is built of 60 quasisymmetric CP dimers (A and B) stabilized by domain swapping, and only the CP-A N-terminal basic region interacts with the packaged nucleic acids. hPBV CP has an α-helical domain with a fold similar to that of fungal partitivirus CP, with many domain insertions in its C-terminal half. In contrast to dsRNA mycoviruses, hPBV has an extracellular life cycle phase like complex reoviruses, which indicates that its own CP probably participates in cell entry. Using an in vitro reversible assembly/disassembly system of hPBV, we isolated tetramers as possible assembly intermediates. We used atomic force microscopy to characterize the biophysical properties of hPBV capsids with different cargos (host nucleic acids or proteins) and found that the CP N-terminal segment not only is involved in nucleic acid interaction/packaging but also modulates the mechanical behavior of the capsid in conjunction with the cargo. IMPORTANCE Despite intensive study, human virus sampling is still sparse, especially for viruses that cause mild or asymptomatic disease. Human picobirnavirus (hPBV) is a double-stranded-RNA virus, broadly dispersed in the human population, but its pathogenicity is uncertain. Here, we report the hPBV structure derived from cryo-electron microscopy (cryo-EM) and reconstruction methods using three capsid protein variants (of different lengths and N-terminal amino acid compositions) that assemble as virus-like particles with distinct properties. The hPBV near-atomic structure reveals a quasisymmetric dimer as the structural subunit and tetramers as possible assembly intermediates that coassemble with nucleic acids. Our structural studies and atomic force microscopy analyses indicate that hPBV capsids are potentially excellent nanocages for gene therapy and targeted drug delivery in humans. (Copyright © 2020 American Society for Microbiology.)
Databáze:	MEDLINE
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