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Interactions between the P6 and P5-1 proteins of southern rice black-streaked dwarf fijivirus in yeast and plant cells

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Abstract

Southern rice black-streaked dwarf virus (SRBSDV) is a recently described member of the genus Fijivirus, family Reoviridae. The roles of the proteins encoded by the SRBSDV genome have rarely been studied. In a yeast two-hybrid (YTH) assay in which SRBSDV P6, a putatively multifunctional protein, was used as bait and an SRBSDV cDNA library was used as prey, there was a strong interaction between the P6 and P5-1 proteins. The interaction was confirmed by bimolecular fluorescence complement (BiFC) assay in plant cells. YTH analysis using truncated mutants showed that the N-terminal region (amino acids 9-231) of P5-1 is necessary for binding P5-1 to P6 and that the N-terminal fragment (amino acids 1-93) of P6 is necessary for its interaction with P5-1. SRBSDV P5-1 formed granules positioned at the cell periphery in Nicotiana benthamiana leaves; P6 was present in both the cytoplasm and the nucleus and formed punctate bodies associated with the cell periphery. Immunogold labeling showed that both P6 and P5-1 localized within viroplasms in infected cells of rice plants. These results suggest that the interaction between P5-1 and P6 of SRBSDV may be involved in the formation of viroplasms.

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Acknowledgments

This work was funded by the China 973 Program (2006CB708209), 863 program (2007AA10Z414), National Science and Technology Support Program (2012BAD19B03), the International Science and Technology Cooperation Project (2007DFB30350), the Special Fund for Agro-scientific Research in the Public Interest of China (201003031), the Zhejiang Provincial Science and Technology Project (2010C12027), and the Zhejiang Provincial Foundation for Natural Science (Z305165 and Y3090657). We thank Professor M. J. Adams, Rothamsted Research, Harpenden, UK, for help in correcting the English of the manuscript.

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Correspondence to Heng-Mu Zhang or Jian-Ping Chen.

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J. Li and J. Xue contributed equally to this work.

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Li, J., Xue, J., Zhang, HM. et al. Interactions between the P6 and P5-1 proteins of southern rice black-streaked dwarf fijivirus in yeast and plant cells. Arch Virol 158, 1649–1659 (2013). https://doi.org/10.1007/s00705-013-1660-4

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  • DOI: https://doi.org/10.1007/s00705-013-1660-4

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