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Article

A Replicating Viral Vector Greatly Enhances Accumulation of Helical Virus-Like Particles in Plants

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John Innes Centre, Department of Biochemistry and Metabolism, Norwich Research Park, Norwich NR4 7UH, UK
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Astbury Centre for Structural Molecular Biology, School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UK
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Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90095, USA
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Centre for Research in Agricultural Genomics (CRAG, CSIC-IRTA-UAB-UB), 08193 Cerdanyola del Vallès, Spain
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Consejo Superior de Investigaciones Científicas (CSIC), 08003 Barcelona, Spain
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Diamante srl. Strada Le Grazie, 15, 37134 Verona, Italy
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Piramal Healthcare UK Ltd., Piramal Pharma Solutions, Earls Road, Grangemouth, Stirlingshire FK3 8XG, UK
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Department of Biotechnology, University of Verona, Strada Le Grazie, 15, 37134 Verona, Italy
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Authors to whom correspondence should be addressed.
Academic Editor: Henryk Czosnek
Viruses 2021, 13(5), 885; https://doi.org/10.3390/v13050885
Received: 6 April 2021 / Revised: 6 May 2021 / Accepted: 7 May 2021 / Published: 11 May 2021
(This article belongs to the Section Viruses of Plants, Fungi and Protozoa)
The production of plant helical virus-like particles (VLPs) via plant-based expression has been problematic with previous studies suggesting that an RNA scaffold may be necessary for their efficient production. To examine this, we compared the accumulation of VLPs from two potexviruses, papaya mosaic virus and alternanthera mosaic virus (AltMV), when the coat proteins were expressed from a replicating potato virus X- based vector (pEff) and a non-replicating vector (pEAQ-HT). Significantly greater quantities of VLPs could be purified when pEff was used. The pEff system was also very efficient at producing VLPs of helical viruses from different virus families. Examination of the RNA content of AltMV and tobacco mosaic virus VLPs produced from pEff revealed the presence of vector-derived RNA sequences, suggesting that the replicating RNA acts as a scaffold for VLP assembly. Cryo-EM analysis of the AltMV VLPs showed they had a structure very similar to that of authentic potexvirus particles. Thus, we conclude that vectors generating replicating forms of RNA, such as pEff, are very efficient for producing helical VLPs. View Full-Text
Keywords: virus-like particle; overexpression; filamentous; rod-shaped; pEAQ-HT; pEff; Potexvirus; Potyviridae; high-resolution cryo-EM virus-like particle; overexpression; filamentous; rod-shaped; pEAQ-HT; pEff; Potexvirus; Potyviridae; high-resolution cryo-EM
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MDPI and ACS Style

Thuenemann, E.C.; Byrne, M.J.; Peyret, H.; Saunders, K.; Castells-Graells, R.; Ferriol, I.; Santoni, M.; Steele, J.F.C.; Ranson, N.A.; Avesani, L.; Lopez-Moya, J.J.; Lomonossoff, G.P. A Replicating Viral Vector Greatly Enhances Accumulation of Helical Virus-Like Particles in Plants. Viruses 2021, 13, 885. https://doi.org/10.3390/v13050885

AMA Style

Thuenemann EC, Byrne MJ, Peyret H, Saunders K, Castells-Graells R, Ferriol I, Santoni M, Steele JFC, Ranson NA, Avesani L, Lopez-Moya JJ, Lomonossoff GP. A Replicating Viral Vector Greatly Enhances Accumulation of Helical Virus-Like Particles in Plants. Viruses. 2021; 13(5):885. https://doi.org/10.3390/v13050885

Chicago/Turabian Style

Thuenemann, Eva C., Matthew J. Byrne, Hadrien Peyret, Keith Saunders, Roger Castells-Graells, Inmaculada Ferriol, Mattia Santoni, John F.C. Steele, Neil A. Ranson, Linda Avesani, Juan J. Lopez-Moya, and George P. Lomonossoff 2021. "A Replicating Viral Vector Greatly Enhances Accumulation of Helical Virus-Like Particles in Plants" Viruses 13, no. 5: 885. https://doi.org/10.3390/v13050885

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