Analyses of pea necrotic yellow dwarf virus-encoded proteins
Pea necrotic yellow dwarf virus (PNYDV) is a multipartite, circular, single-stranded DNA plant virus. PNYDV encodes eight proteins and the function of three of which remains unknown—U1, U2, and U4. PNYDV proteins cellular localization was analyzed by GFP tagging and bimolecular fluorescence complementation (BiFC) studies. The interactions of all eight PNYDV proteins were tested pairwise in planta (36 combinations in total). Seven interactions were identified and two (M-Rep with CP and MP with U4) were characterized further. MP and U4 complexes appeared as vesicle-like spots and were localized at the nuclear envelope and cell periphery. These vesicle-like spots were associated with the endoplasmatic reticulum. In addition, a nuclear localization signal (NLS) was mapped for U1, and a mutated U1 with NLS disrupted localized at plasmodesmata and therefore might also have a role in movement. Taken together, this study provides evidence for previously undescribed nanovirus protein–protein interactions and their cellular localization with novel findings not only for those proteins with unknown function, but also for characterized proteins such as the CP.
KeywordsNanoviridae Nanovirus Pea necrotic yellow dwarf virus Bimolecular fluorescence complementation (BiFC)
This work was funded by Grants of the Deutsche Forschungsgemeinschaft (SFB796 subproject AP2). The authors would like to thank the gardener Christine Hösl for taking care of experimental plants. We are very grateful to Bruno Gronenborn, Tatjana Kleinow (University of Stuttgart, Germany), and Frederik Börnke (IGZ—Leibniz Institute of Vegetable and Ornamental Crops, Großbeeren, Germany) for providing plasmids and support on the BiFC analysis. We would like to acknowledge the Optical Imaging Centre Erlangen (OICE). We are also grateful to Jeremy R. Thompson (Cornell University, Ithaca, USA) for critically reading of the manuscript and helpful discussions.
BK. and S.K. wrote the manuscript, B.K., I.S., S.K., and E.G. performed research, and B.K. and S.K. designed the study.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Research involving human participants and/or animals
This article does not contain any studies with human participants or animals performed by any of the authors.
Informed consent was obtained from all individual participants included in the study.
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