Investigating the Viral Suppressor HC-Pro Inhibiting Small RNA Methylation through Functional Comparison of HEN1 in Angiosperm and Bryophyte
In plants, HEN1-facilitated methylation at 3′ end ribose is a critical step of small-RNA (sRNA) biogenesis. A mutant of well-studied Arabidopsis HEN1 (AtHEN1), <i>hen1-1</i>, showed a defective developmental phenotype, indicating the importance of sRNA methylation. Moreover, <i>Mar...
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MDPI AG
2021-09-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/13/9/1837 |
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| author | Neda Sanobar Pin-Chun Lin Zhao-Jun Pan Ru-Ying Fang Veny Tjita Fang-Fang Chen Hao-Ching Wang Huang-Lung Tsai Shu-Hsing Wu Tang-Long Shen Yan-Huey Chen Shih-Shun Lin |
| author_facet | Neda Sanobar Pin-Chun Lin Zhao-Jun Pan Ru-Ying Fang Veny Tjita Fang-Fang Chen Hao-Ching Wang Huang-Lung Tsai Shu-Hsing Wu Tang-Long Shen Yan-Huey Chen Shih-Shun Lin |
| author_sort | Neda Sanobar |
| collection | DOAJ |
| description | In plants, HEN1-facilitated methylation at 3′ end ribose is a critical step of small-RNA (sRNA) biogenesis. A mutant of well-studied Arabidopsis HEN1 (AtHEN1), <i>hen1-1</i>, showed a defective developmental phenotype, indicating the importance of sRNA methylation. Moreover, <i>Marchantia polymorpha</i> has been identified to have a <i>HEN1</i> ortholog gene (Mp<i>HEN1</i>); however, its function remained unfathomed. Our in vivo and in vitro data have shown MpHEN1 activity being comparable with AtHEN1, and their substrate specificity towards duplex microRNA (miRNA) remained consistent. Furthermore, the phylogenetic tree and multiple alignment highlighted the conserved molecular evolution of the HEN1 family in plants. The P1/HC-Pro of the turnip mosaic virus (TuMV) is a known RNA silencing suppressor and inhibits HEN1 methylation of sRNAs. Here, we report that the HC-Pro physically binds with AtHEN1 through FRNK motif, inhibiting HEN1’s methylation activity. Moreover, the in vitro EMSA data indicates GST-HC-Pro of TuMV lacks sRNA duplex-binding ability. Surprisingly, the HC-Pro also inhibits MpHEN1 activity in a dosage-dependent manner, suggesting the possibility of interaction between HC-Pro and MpHEN1 as well. Further investigations on understanding interaction mechanisms of HEN1 and various HC-Pros can advance the knowledge of viral suppressors. |
| first_indexed | 2024-03-10T07:08:21Z |
| format | Article |
| id | doaj.art-198b0607a73045e18471aff3be2ccab1 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T07:08:21Z |
| publishDate | 2021-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-198b0607a73045e18471aff3be2ccab12023-11-22T15:38:43ZengMDPI AGViruses1999-49152021-09-01139183710.3390/v13091837Investigating the Viral Suppressor HC-Pro Inhibiting Small RNA Methylation through Functional Comparison of HEN1 in Angiosperm and BryophyteNeda Sanobar0Pin-Chun Lin1Zhao-Jun Pan2Ru-Ying Fang3Veny Tjita4Fang-Fang Chen5Hao-Ching Wang6Huang-Lung Tsai7Shu-Hsing Wu8Tang-Long Shen9Yan-Huey Chen10Shih-Shun Lin11Institute of Biotechnology, National Taiwan University, Taipei 106, TaiwanInstitute of Biotechnology, National Taiwan University, Taipei 106, TaiwanInstitute of Biotechnology, National Taiwan University, Taipei 106, TaiwanInstitute of Biotechnology, National Taiwan University, Taipei 106, TaiwanInstitute of Biotechnology, National Taiwan University, Taipei 106, TaiwanInstitute of Biotechnology, National Taiwan University, Taipei 106, TaiwanThe PhD Program for Translational Medicine, College of Medical Science and Technology, Taipei Medical University and Academia Sinica, Taipei 115, TaiwanInstitute of Plant and Microbial Biology, Academia Sinica, Taipei 115, TaiwanInstitute of Plant and Microbial Biology, Academia Sinica, Taipei 115, TaiwanCenter of Biotechnology, National Taiwan University, Taipei 106, TaiwanInstitute of Biotechnology, National Taiwan University, Taipei 106, TaiwanInstitute of Biotechnology, National Taiwan University, Taipei 106, TaiwanIn plants, HEN1-facilitated methylation at 3′ end ribose is a critical step of small-RNA (sRNA) biogenesis. A mutant of well-studied Arabidopsis HEN1 (AtHEN1), <i>hen1-1</i>, showed a defective developmental phenotype, indicating the importance of sRNA methylation. Moreover, <i>Marchantia polymorpha</i> has been identified to have a <i>HEN1</i> ortholog gene (Mp<i>HEN1</i>); however, its function remained unfathomed. Our in vivo and in vitro data have shown MpHEN1 activity being comparable with AtHEN1, and their substrate specificity towards duplex microRNA (miRNA) remained consistent. Furthermore, the phylogenetic tree and multiple alignment highlighted the conserved molecular evolution of the HEN1 family in plants. The P1/HC-Pro of the turnip mosaic virus (TuMV) is a known RNA silencing suppressor and inhibits HEN1 methylation of sRNAs. Here, we report that the HC-Pro physically binds with AtHEN1 through FRNK motif, inhibiting HEN1’s methylation activity. Moreover, the in vitro EMSA data indicates GST-HC-Pro of TuMV lacks sRNA duplex-binding ability. Surprisingly, the HC-Pro also inhibits MpHEN1 activity in a dosage-dependent manner, suggesting the possibility of interaction between HC-Pro and MpHEN1 as well. Further investigations on understanding interaction mechanisms of HEN1 and various HC-Pros can advance the knowledge of viral suppressors.https://www.mdpi.com/1999-4915/13/9/1837sRNAHEN1methylationviral suppressorsHEN1-HC-Pro interaction |
| spellingShingle | Neda Sanobar Pin-Chun Lin Zhao-Jun Pan Ru-Ying Fang Veny Tjita Fang-Fang Chen Hao-Ching Wang Huang-Lung Tsai Shu-Hsing Wu Tang-Long Shen Yan-Huey Chen Shih-Shun Lin Investigating the Viral Suppressor HC-Pro Inhibiting Small RNA Methylation through Functional Comparison of HEN1 in Angiosperm and Bryophyte Viruses sRNA HEN1 methylation viral suppressors HEN1-HC-Pro interaction |
| title | Investigating the Viral Suppressor HC-Pro Inhibiting Small RNA Methylation through Functional Comparison of HEN1 in Angiosperm and Bryophyte |
| title_full | Investigating the Viral Suppressor HC-Pro Inhibiting Small RNA Methylation through Functional Comparison of HEN1 in Angiosperm and Bryophyte |
| title_fullStr | Investigating the Viral Suppressor HC-Pro Inhibiting Small RNA Methylation through Functional Comparison of HEN1 in Angiosperm and Bryophyte |
| title_full_unstemmed | Investigating the Viral Suppressor HC-Pro Inhibiting Small RNA Methylation through Functional Comparison of HEN1 in Angiosperm and Bryophyte |
| title_short | Investigating the Viral Suppressor HC-Pro Inhibiting Small RNA Methylation through Functional Comparison of HEN1 in Angiosperm and Bryophyte |
| title_sort | investigating the viral suppressor hc pro inhibiting small rna methylation through functional comparison of hen1 in angiosperm and bryophyte |
| topic | sRNA HEN1 methylation viral suppressors HEN1-HC-Pro interaction |
| url | https://www.mdpi.com/1999-4915/13/9/1837 |
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