Turnip Mosaic Virus Coat Protein Deletion Mutants Allow Defining Dispensable Protein Domains for ‘in Planta’ eVLP Formation
The involvement of different structural domains of the coat protein (CP) of turnip mosaic virus, a potyvirus, in establishing and/or maintaining particle assembly was analyzed through deletion mutants of the protein. In order to identify exclusively those domains involved in protein–protein interact...
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MDPI AG
2020-06-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/12/6/661 |
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| author | Carmen Yuste-Calvo Pablo Ibort Flora Sánchez Fernando Ponz |
| author_facet | Carmen Yuste-Calvo Pablo Ibort Flora Sánchez Fernando Ponz |
| author_sort | Carmen Yuste-Calvo |
| collection | DOAJ |
| description | The involvement of different structural domains of the coat protein (CP) of turnip mosaic virus, a potyvirus, in establishing and/or maintaining particle assembly was analyzed through deletion mutants of the protein. In order to identify exclusively those domains involved in protein–protein interactions within the particle, the analysis was performed by agroinfiltration “in planta”, followed by the assessment of CP accumulation in leaves and the assembly of virus-like particles lacking nucleic acids, also known as empty virus-like particles (eVLP). Thus, the interactions involving viral RNA could be excluded. It was found that deletions precluding eVLP assembly did not allow for protein accumulation either, probably indicating that non-assembled CP protein was degraded in the plant leaves. Deletions involving the CP structural core were incompatible with particle assembly. On the N-terminal domain, only the deletion avoiding the subdomain involved in interactions with other CP subunits was incorporated into eVLPs. The C-terminal domain was shown to be more permissive to deletions. Assembled eVLPs were found for mutants, eliminating the whole domain. The C-terminal domain mutants were unusually long, suggesting some role of the domain in the regulation of particle length. The identification of the CP domains responsible for eVLP formation will allow for new approaches to protein stretch replacement with peptides or proteins of nanobiotechnological interest. Finally, specific cases of application are considered. |
| first_indexed | 2024-03-10T19:02:45Z |
| format | Article |
| id | doaj.art-f69c6a9214ed4f42bcb608ae2af2cc01 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T19:02:45Z |
| publishDate | 2020-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-f69c6a9214ed4f42bcb608ae2af2cc012023-11-20T04:20:24ZengMDPI AGViruses1999-49152020-06-0112666110.3390/v12060661Turnip Mosaic Virus Coat Protein Deletion Mutants Allow Defining Dispensable Protein Domains for ‘in Planta’ eVLP FormationCarmen Yuste-Calvo0Pablo Ibort1Flora Sánchez2Fernando Ponz3Centro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid-Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (CBGP, UPM-INIA), Campus Montegancedo, Autopista M-40, km 38, Pozuelo de Alarcón, 28223 Madrid, SpainCentro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid-Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (CBGP, UPM-INIA), Campus Montegancedo, Autopista M-40, km 38, Pozuelo de Alarcón, 28223 Madrid, SpainCentro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid-Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (CBGP, UPM-INIA), Campus Montegancedo, Autopista M-40, km 38, Pozuelo de Alarcón, 28223 Madrid, SpainCentro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid-Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (CBGP, UPM-INIA), Campus Montegancedo, Autopista M-40, km 38, Pozuelo de Alarcón, 28223 Madrid, SpainThe involvement of different structural domains of the coat protein (CP) of turnip mosaic virus, a potyvirus, in establishing and/or maintaining particle assembly was analyzed through deletion mutants of the protein. In order to identify exclusively those domains involved in protein–protein interactions within the particle, the analysis was performed by agroinfiltration “in planta”, followed by the assessment of CP accumulation in leaves and the assembly of virus-like particles lacking nucleic acids, also known as empty virus-like particles (eVLP). Thus, the interactions involving viral RNA could be excluded. It was found that deletions precluding eVLP assembly did not allow for protein accumulation either, probably indicating that non-assembled CP protein was degraded in the plant leaves. Deletions involving the CP structural core were incompatible with particle assembly. On the N-terminal domain, only the deletion avoiding the subdomain involved in interactions with other CP subunits was incorporated into eVLPs. The C-terminal domain was shown to be more permissive to deletions. Assembled eVLPs were found for mutants, eliminating the whole domain. The C-terminal domain mutants were unusually long, suggesting some role of the domain in the regulation of particle length. The identification of the CP domains responsible for eVLP formation will allow for new approaches to protein stretch replacement with peptides or proteins of nanobiotechnological interest. Finally, specific cases of application are considered.https://www.mdpi.com/1999-4915/12/6/661VNPsassemblyeVLPsmutantdeletionpotyvirus |
| spellingShingle | Carmen Yuste-Calvo Pablo Ibort Flora Sánchez Fernando Ponz Turnip Mosaic Virus Coat Protein Deletion Mutants Allow Defining Dispensable Protein Domains for ‘in Planta’ eVLP Formation Viruses VNPs assembly eVLPs mutant deletion potyvirus |
| title | Turnip Mosaic Virus Coat Protein Deletion Mutants Allow Defining Dispensable Protein Domains for ‘in Planta’ eVLP Formation |
| title_full | Turnip Mosaic Virus Coat Protein Deletion Mutants Allow Defining Dispensable Protein Domains for ‘in Planta’ eVLP Formation |
| title_fullStr | Turnip Mosaic Virus Coat Protein Deletion Mutants Allow Defining Dispensable Protein Domains for ‘in Planta’ eVLP Formation |
| title_full_unstemmed | Turnip Mosaic Virus Coat Protein Deletion Mutants Allow Defining Dispensable Protein Domains for ‘in Planta’ eVLP Formation |
| title_short | Turnip Mosaic Virus Coat Protein Deletion Mutants Allow Defining Dispensable Protein Domains for ‘in Planta’ eVLP Formation |
| title_sort | turnip mosaic virus coat protein deletion mutants allow defining dispensable protein domains for in planta evlp formation |
| topic | VNPs assembly eVLPs mutant deletion potyvirus |
| url | https://www.mdpi.com/1999-4915/12/6/661 |
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