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A non-structural protein encoded by Rice Dwarf Virus targets to the nucleus and chloroplast and inhibits local RNA silencing

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Abstract

RNA silencing is a potent antiviral mechanism in plants and animals. As a counter-defense, many viruses studied to date encode one or more viral suppressors of RNA silencing (VSR). In the latter case, how different VSRs encoded by a virus function in silencing remains to be fully understood. We previously showed that the nonstructural protein Pns10 of a Phytoreovirus, Rice dwarf virus (RDV), functions as a VSR. Here we present evidence that another nonstructural protein, Pns11, also functions as a VSR. While Pns10 was localized in the cytoplasm, Pns11 was localized both in the nucleus and chloroplasts. Pns11 has two bipartite nuclear localization signals (NLSs), which were required for nuclear as well as chloroplastic localization. The NLSs were also required for the silencing activities of Pns11. This is the first report that multiple VSRs encoded by a virus are localized in different subcellular compartments, and that a viral protein can be targeted to both the nucleus and chloroplast. These findings may have broad significance in studying the subcellular targeting of VSRs and other viral proteins in viral-host interactions.

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Acknowledgements

We thank Prof.Biao Dingfor critical comments and substantial reading the manuscript, Prof. Shouwei Ding for valuable suggestions and providing plasmid pCamBia1300-Tav2b, Prof. David Baulcombe for providing 16c seeds. We also like to thank Dr.s Yiming Bao, Bo Ren, Qiuting Ren for their assistance. This work was supported by grants from CARS-01-06 to X.H.X., Transgenic Research Program (2016ZX08010001) and the National Natural Science Foundation of China (31530062) to Y.L.

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Authors and Affiliations

  1. The State Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, 100871, China

    Feng Gao, Shanshan Zhao, Chunhong Wei, Wei Hong & Yi Li

  2. Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, Maryland, 20742, USA

    Feng Gao

  3. College of Plant Protection, Fujian Agriculture & Forestry University, Fuzhou, 350002, China

    Shanshan Zhao

  4. College of Life Sciences, Nankai University, Tianjin, 300071, China

    Shuzhen Men

  5. Department of Plant Protection, Northwestern Agriculture and Forestry University, Yangling, 712100, China

    Zhensheng Kang

  6. College of Agriculture, Zhejiang University, Hangzhou, 310029, China

    Jian Hong

  7. The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, 310006, China

    Wei Hong

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  1. Feng Gao
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  2. Shanshan Zhao
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  3. Shuzhen Men
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Correspondence to Wei Hong or Yi Li.

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Supporting Information

Figure S1 Schematic of two products encoded by the full-length segment S11 of RDV. Pns11 was used in this study.

Figure S2 Subcellular localization of transiently expressed Pns11-GFP in epidermal cells of N. benthamiana.

Figure S3 Pns11-GFP and various Pns11-GFP mutants were transiently expressed in BY2 cells.

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Gao, F., Zhao, S., Men, S. et al. A non-structural protein encoded by Rice Dwarf Virus targets to the nucleus and chloroplast and inhibits local RNA silencing. Sci. China Life Sci. 63, 1703–1713 (2020). https://doi.org/10.1007/s11427-019-1648-3

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