Cauliflower mosaic virus Transcriptome Reveals a Complex Alternative Splicing Pattern.
The plant pararetrovirus Cauliflower mosaic virus (CaMV) uses alternative splicing to generate several isoforms from its polycistronic pregenomic 35S RNA. This pro-cess has been shown to be essential for infectivity. Previous works have identified four splice donor sites and a single splice acceptor...
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| Format: | Article |
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Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4498817?pdf=render |
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| author | Clément Bouton Angèle Geldreich Laëtitia Ramel Lyubov A Ryabova Maria Dimitrova Mario Keller |
| author_facet | Clément Bouton Angèle Geldreich Laëtitia Ramel Lyubov A Ryabova Maria Dimitrova Mario Keller |
| author_sort | Clément Bouton |
| collection | DOAJ |
| description | The plant pararetrovirus Cauliflower mosaic virus (CaMV) uses alternative splicing to generate several isoforms from its polycistronic pregenomic 35S RNA. This pro-cess has been shown to be essential for infectivity. Previous works have identified four splice donor sites and a single splice acceptor site in the 35S RNA 5' region and suggested that the main role of CaMV splicing is to downregulate expression of open reading frames (ORFs) I and II. In this study, we show that alternative splicing is a conserved process among CaMV isolates. In Cabb B-JI and Cabb-S isolates, splicing frequently leads to different fusion between ORFs, particularly between ORF I and II. The corresponding P1P2 fusion proteins expressed in E. coli interact with viral proteins P2 and P3 in vitro. However, they are detected neither during infection nor upon transient expression in planta, which suggests rapid degradation after synthesis and no important biological role in the CaMV infectious cycle. To gain a better understanding of the functional relevance of 35S RNA alternative splicing in CaMV infectivity, we inactivated the previously described splice sites. All the splicing mutants were as pathogenic as the corresponding wild-type isolate. Through RT-PCR-based analysis we demonstrate that CaMV 35S RNA exhibits a complex splicing pattern, as we identify new splice donor and acceptor sites whose selection leads to more than thirteen 35S RNA isoforms in infected turnip plants. Inactivating splice donor or acceptor sites is not lethal for the virus, since disrupted sites are systematically rescued by the activation of cryptic and/or seldom used splice sites. Taken together, our data depict a conserved, complex and flexible process, involving multiple sites, that ensures splicing of 35S RNA. |
| first_indexed | 2024-12-20T08:56:28Z |
| format | Article |
| id | doaj.art-5c171de50f054430b82a6e924e91298c |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-20T08:56:28Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-5c171de50f054430b82a6e924e91298c2022-12-21T19:45:59ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01107e013266510.1371/journal.pone.0132665Cauliflower mosaic virus Transcriptome Reveals a Complex Alternative Splicing Pattern.Clément BoutonAngèle GeldreichLaëtitia RamelLyubov A RyabovaMaria DimitrovaMario KellerThe plant pararetrovirus Cauliflower mosaic virus (CaMV) uses alternative splicing to generate several isoforms from its polycistronic pregenomic 35S RNA. This pro-cess has been shown to be essential for infectivity. Previous works have identified four splice donor sites and a single splice acceptor site in the 35S RNA 5' region and suggested that the main role of CaMV splicing is to downregulate expression of open reading frames (ORFs) I and II. In this study, we show that alternative splicing is a conserved process among CaMV isolates. In Cabb B-JI and Cabb-S isolates, splicing frequently leads to different fusion between ORFs, particularly between ORF I and II. The corresponding P1P2 fusion proteins expressed in E. coli interact with viral proteins P2 and P3 in vitro. However, they are detected neither during infection nor upon transient expression in planta, which suggests rapid degradation after synthesis and no important biological role in the CaMV infectious cycle. To gain a better understanding of the functional relevance of 35S RNA alternative splicing in CaMV infectivity, we inactivated the previously described splice sites. All the splicing mutants were as pathogenic as the corresponding wild-type isolate. Through RT-PCR-based analysis we demonstrate that CaMV 35S RNA exhibits a complex splicing pattern, as we identify new splice donor and acceptor sites whose selection leads to more than thirteen 35S RNA isoforms in infected turnip plants. Inactivating splice donor or acceptor sites is not lethal for the virus, since disrupted sites are systematically rescued by the activation of cryptic and/or seldom used splice sites. Taken together, our data depict a conserved, complex and flexible process, involving multiple sites, that ensures splicing of 35S RNA.http://europepmc.org/articles/PMC4498817?pdf=render |
| spellingShingle | Clément Bouton Angèle Geldreich Laëtitia Ramel Lyubov A Ryabova Maria Dimitrova Mario Keller Cauliflower mosaic virus Transcriptome Reveals a Complex Alternative Splicing Pattern. PLoS ONE |
| title | Cauliflower mosaic virus Transcriptome Reveals a Complex Alternative Splicing Pattern. |
| title_full | Cauliflower mosaic virus Transcriptome Reveals a Complex Alternative Splicing Pattern. |
| title_fullStr | Cauliflower mosaic virus Transcriptome Reveals a Complex Alternative Splicing Pattern. |
| title_full_unstemmed | Cauliflower mosaic virus Transcriptome Reveals a Complex Alternative Splicing Pattern. |
| title_short | Cauliflower mosaic virus Transcriptome Reveals a Complex Alternative Splicing Pattern. |
| title_sort | cauliflower mosaic virus transcriptome reveals a complex alternative splicing pattern |
| url | http://europepmc.org/articles/PMC4498817?pdf=render |
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