<i>Trans</i>-Activator Binding Site Context in RCNMV Modulates Subgenomic mRNA Transcription
Many positive-sense RNA viruses transcribe subgenomic (sg) mRNAs during infections that template the translation of a subset of viral proteins. Red clover necrotic mosaic virus (RCNMV) expresses its capsid protein through the transcription of a sg mRNA from RNA1 genome segment. This transcription ev...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-11-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/13/11/2252 |
| _version_ | 1797508189031235584 |
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| author | Jennifer S. H. Im Laura R. Newburn Gregory Kent K. Andrew White |
| author_facet | Jennifer S. H. Im Laura R. Newburn Gregory Kent K. Andrew White |
| author_sort | Jennifer S. H. Im |
| collection | DOAJ |
| description | Many positive-sense RNA viruses transcribe subgenomic (sg) mRNAs during infections that template the translation of a subset of viral proteins. Red clover necrotic mosaic virus (RCNMV) expresses its capsid protein through the transcription of a sg mRNA from RNA1 genome segment. This transcription event is activated by an RNA structure formed by base pairing between a <i>trans</i>-activator (TA) in RNA2 and a <i>trans</i>-activator binding site (TABS) in RNA1. In this study, the impact of the structural context of the TABS in RNA1 on the TA–TABS interaction and sg mRNA transcription was investigated using in vitro and in vivo approaches. The results (i) generated RNA secondary structure models for the TA and TABS, (ii) revealed that the TABS is partially base paired with proximal upstream sequences, which limits TA access, (iii) demonstrated that the aforementioned intra-RNA1 base pairing involving the TABS modulates the TA–TABS interaction in vitro and sg mRNA levels during infections, and (iv) revealed that the TABS in RNA1 can be modified to mediate sg mRNA transcription in a TA-independent manner. These findings advance our understanding of transcriptional regulation in RCNMV and provide novel insights into the origin of the TA–TABS interaction. |
| first_indexed | 2024-03-10T04:58:46Z |
| format | Article |
| id | doaj.art-02225d4429214cc2808e812a4d30513b |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T04:58:46Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-02225d4429214cc2808e812a4d30513b2023-11-23T01:57:35ZengMDPI AGViruses1999-49152021-11-011311225210.3390/v13112252<i>Trans</i>-Activator Binding Site Context in RCNMV Modulates Subgenomic mRNA TranscriptionJennifer S. H. Im0Laura R. Newburn1Gregory Kent2K. Andrew White3Department of Biology, York University, Toronto, ON M3J 1P3, CanadaDepartment of Biology, York University, Toronto, ON M3J 1P3, CanadaDepartment of Biology, York University, Toronto, ON M3J 1P3, CanadaDepartment of Biology, York University, Toronto, ON M3J 1P3, CanadaMany positive-sense RNA viruses transcribe subgenomic (sg) mRNAs during infections that template the translation of a subset of viral proteins. Red clover necrotic mosaic virus (RCNMV) expresses its capsid protein through the transcription of a sg mRNA from RNA1 genome segment. This transcription event is activated by an RNA structure formed by base pairing between a <i>trans</i>-activator (TA) in RNA2 and a <i>trans</i>-activator binding site (TABS) in RNA1. In this study, the impact of the structural context of the TABS in RNA1 on the TA–TABS interaction and sg mRNA transcription was investigated using in vitro and in vivo approaches. The results (i) generated RNA secondary structure models for the TA and TABS, (ii) revealed that the TABS is partially base paired with proximal upstream sequences, which limits TA access, (iii) demonstrated that the aforementioned intra-RNA1 base pairing involving the TABS modulates the TA–TABS interaction in vitro and sg mRNA levels during infections, and (iv) revealed that the TABS in RNA1 can be modified to mediate sg mRNA transcription in a TA-independent manner. These findings advance our understanding of transcriptional regulation in RCNMV and provide novel insights into the origin of the TA–TABS interaction.https://www.mdpi.com/1999-4915/13/11/2252RNA virusplant virus<i>Tombusviridae</i>tombusvirusdianthovirusRNA–RNA |
| spellingShingle | Jennifer S. H. Im Laura R. Newburn Gregory Kent K. Andrew White <i>Trans</i>-Activator Binding Site Context in RCNMV Modulates Subgenomic mRNA Transcription Viruses RNA virus plant virus <i>Tombusviridae</i> tombusvirus dianthovirus RNA–RNA |
| title | <i>Trans</i>-Activator Binding Site Context in RCNMV Modulates Subgenomic mRNA Transcription |
| title_full | <i>Trans</i>-Activator Binding Site Context in RCNMV Modulates Subgenomic mRNA Transcription |
| title_fullStr | <i>Trans</i>-Activator Binding Site Context in RCNMV Modulates Subgenomic mRNA Transcription |
| title_full_unstemmed | <i>Trans</i>-Activator Binding Site Context in RCNMV Modulates Subgenomic mRNA Transcription |
| title_short | <i>Trans</i>-Activator Binding Site Context in RCNMV Modulates Subgenomic mRNA Transcription |
| title_sort | i trans i activator binding site context in rcnmv modulates subgenomic mrna transcription |
| topic | RNA virus plant virus <i>Tombusviridae</i> tombusvirus dianthovirus RNA–RNA |
| url | https://www.mdpi.com/1999-4915/13/11/2252 |
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