Interaction of Mal de Río Cuarto virus (Fijivirus genus) proteins and identification of putative factors determining viroplasm formation and decay
Mal de Río Cuarto virus (MRCV) is a member of the Fijivirus genus, within the Reoviridae family, that repli-cates and assembles in cytoplasmic inclusion bodies called viroplasms. In this study, we investigatedinteractions between ten MRCV proteins by yeast two-hybrid (Y2H) assays and identified inte...
| Autores principales: | , , , |
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| Formato: | info:ar-repo/semantics/artículo |
| Lenguaje: | Inglés |
| Publicado: |
2017
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/954 http://www.sciencedirect.com/science/article/pii/S0168170216307195?via%3Dihub https://doi.org/10.1016/j.virusres.2017.01.002 |
| Sumario: | Mal de Río Cuarto virus (MRCV) is a member of the Fijivirus genus, within the Reoviridae family, that repli-cates and assembles in cytoplasmic inclusion bodies called viroplasms. In this study, we investigatedinteractions between ten MRCV proteins by yeast two-hybrid (Y2H) assays and identified interactionsof non-structural proteins P6/P6, P9-2/P9-2 and P6/P9-1. P9-1 and P6 are the major and minor compo-nents of the viroplasms respectively, whereas P9-2 is an N-glycosylated membrane protein of unknownfunction. Interactions involving P6 and P9-1 were confirmed by bimolecular fluorescence complementa-tion (BiFC) in rice protoplasts. We demonstrated that a region including a predicted coiled-coil domainwithin the C-terminal moiety of P6 was necessary for P6/P6 and P6/P9-1 interactions. In turn, a shortC-terminal arm was necessary for the previously reported P9-1 self-interaction. Transient expression ofthese proteins by agroinfiltration of Nicotiana benthamiana leaves showed very low accumulation levelsand further in silico analyses allowed us to identify conserved PEST degradation sequences [rich in pro-line (P), glutamic acid (E), serine (S), and threonine (T)] within P6 and P9-1. The removal of these PESTsequences resulted in a significant increase of the accumulation of both proteins. |
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