title: Detection of plum pox potyviral protein-protein interactions in planta using an optimized mRFP-based bimolecular fluorescence complementation system.
abstract: In previous studies, protein interaction maps of different potyviruses have been generated using yeast two-hybrid (YTH) systems, and these maps have demonstrated a high diversity of interactions of potyviral proteins. Using an optimized bimolecular fluorescence complementation (BiFC) system, a complete interaction matrix for proteins of a potyvirus was developed for the first time under in planta conditions with ten proteins from plum pox virus (PPV). In total, 52 of 100 possible interactions were detected, including the self-interactions of CI, 6K2, VPg, NIa-Pro, NIb and CP, which is more interactions than have ever been detected for any other potyvirus in a YTH approach. Moreover, the BiFC system was shown to be able to localize the protein interactions, which was typified for the protein self-interactions indicated above. Additionally, experiments were carried out with the P3N-PIPO protein, revealing an interaction with CI but not with CP and supporting the involvement of P3N-PIPO in the cell-to-cell movement of potyviruses. No self-interaction of the PPV helper component-proteinase (HC-Pro) was detected using BiFC in planta. Therefore, additional experiments with turnip mosaic virus (TuMV) HC-Pro, PPV_HC-Pro and their mutants were conducted. The self-interaction of TuMV_HCpro, as recently demonstrated, and the self-interaction of the TuMV_ and PPV_HC-Pro mutants were shown by BiFC in planta, indicating that HC-Pro self-interactions may be species-specific. BiFC is a very useful and reliable method for the detection and localization of protein interactions in planta, thus enabling investigations under more natural conditions than studies in yeast cells.
authors: Gottfried Wilhelm Leibniz University of Hannover, Institute of Plant Diseases and Plant Protection, Herrenhäuser Straße 2, D-30419 Hannover, Germany.
authors: Eva
authors: E
authors: Zilian
authors: Gottfried Wilhelm Leibniz University of Hannover, Institute of Plant Diseases and Plant Protection, Herrenhäuser Straße 2, D-30419 Hannover, Germany.
authors: Edgar
authors: E
authors: Maiss
mesh_terms: D003165:Complement System Proteins; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism; D005453:Fluorescence; D054884:Host-Pathogen Interactions; D008164:Luminescent Proteins / Q000378:metabolism; D009154:Mutation; D010957:Plasmids; D017801:Plum Pox Virus / Q000235:genetics* / Q000378:metabolism; D025941:Protein Interaction Mapping / Q000379:methods; D060066:Protein Interaction Maps; D014026:Nicotiana / Q000166:cytology / Q000821:virology*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D000097573:Red Fluorescent Protein
chemical_list: D003165:Complement System Proteins; D003546:Cysteine Endopeptidases; D008164:Luminescent Proteins; D014764:Viral Proteins; D000097573:Red Fluorescent Protein; C072775:HC-Pro protein, potyvirus