A Replicating Viral Vector Greatly Enhances Accumulation of Helical Virus-Like Particles in Plants
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 mosai...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-05-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/13/5/885 |
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| author | Eva C. Thuenemann Matthew J. Byrne Hadrien Peyret Keith Saunders Roger Castells-Graells Inmaculada Ferriol Mattia Santoni John F. C. Steele Neil A. Ranson Linda Avesani Juan Jose Lopez-Moya George P. Lomonossoff |
| author_facet | Eva C. Thuenemann Matthew J. Byrne Hadrien Peyret Keith Saunders Roger Castells-Graells Inmaculada Ferriol Mattia Santoni John F. C. Steele Neil A. Ranson Linda Avesani Juan Jose Lopez-Moya George P. Lomonossoff |
| author_sort | Eva C. Thuenemann |
| collection | DOAJ |
| description | 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-<i>HT</i>). 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. |
| first_indexed | 2024-03-10T11:31:27Z |
| format | Article |
| id | doaj.art-2ba02e7ef9e8407fa6b0d37317efde36 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T11:31:27Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-2ba02e7ef9e8407fa6b0d37317efde362023-11-21T19:13:58ZengMDPI AGViruses1999-49152021-05-0113588510.3390/v13050885A Replicating Viral Vector Greatly Enhances Accumulation of Helical Virus-Like Particles in PlantsEva C. Thuenemann0Matthew J. Byrne1Hadrien Peyret2Keith Saunders3Roger Castells-Graells4Inmaculada Ferriol5Mattia Santoni6John F. C. Steele7Neil A. Ranson8Linda Avesani9Juan Jose Lopez-Moya10George P. Lomonossoff11John Innes Centre, Department of Biochemistry and Metabolism, Norwich Research Park, Norwich NR4 7UH, UKAstbury Centre for Structural Molecular Biology, School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UKJohn Innes Centre, Department of Biochemistry and Metabolism, Norwich Research Park, Norwich NR4 7UH, UKJohn Innes Centre, Department of Biochemistry and Metabolism, Norwich Research Park, Norwich NR4 7UH, UKJohn Innes Centre, Department of Biochemistry and Metabolism, Norwich Research Park, Norwich NR4 7UH, UKCentre for Research in Agricultural Genomics (CRAG, CSIC-IRTA-UAB-UB), 08193 Cerdanyola del Vallès, SpainDiamante srl. Strada Le Grazie, 15, 37134 Verona, ItalyJohn Innes Centre, Department of Biochemistry and Metabolism, Norwich Research Park, Norwich NR4 7UH, UKAstbury Centre for Structural Molecular Biology, School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UKDepartment of Biotechnology, University of Verona, Strada Le Grazie, 15, 37134 Verona, ItalyCentre for Research in Agricultural Genomics (CRAG, CSIC-IRTA-UAB-UB), 08193 Cerdanyola del Vallès, SpainJohn Innes Centre, Department of Biochemistry and Metabolism, Norwich Research Park, Norwich NR4 7UH, UKThe 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-<i>HT</i>). 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.https://www.mdpi.com/1999-4915/13/5/885virus-like particleoverexpressionfilamentousrod-shapedpEAQ-<i>HT</i>pEff |
| spellingShingle | Eva C. Thuenemann Matthew J. Byrne Hadrien Peyret Keith Saunders Roger Castells-Graells Inmaculada Ferriol Mattia Santoni John F. C. Steele Neil A. Ranson Linda Avesani Juan Jose Lopez-Moya George P. Lomonossoff A Replicating Viral Vector Greatly Enhances Accumulation of Helical Virus-Like Particles in Plants Viruses virus-like particle overexpression filamentous rod-shaped pEAQ-<i>HT</i> pEff |
| title | A Replicating Viral Vector Greatly Enhances Accumulation of Helical Virus-Like Particles in Plants |
| title_full | A Replicating Viral Vector Greatly Enhances Accumulation of Helical Virus-Like Particles in Plants |
| title_fullStr | A Replicating Viral Vector Greatly Enhances Accumulation of Helical Virus-Like Particles in Plants |
| title_full_unstemmed | A Replicating Viral Vector Greatly Enhances Accumulation of Helical Virus-Like Particles in Plants |
| title_short | A Replicating Viral Vector Greatly Enhances Accumulation of Helical Virus-Like Particles in Plants |
| title_sort | replicating viral vector greatly enhances accumulation of helical virus like particles in plants |
| topic | virus-like particle overexpression filamentous rod-shaped pEAQ-<i>HT</i> pEff |
| url | https://www.mdpi.com/1999-4915/13/5/885 |
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