title: Fluorescent protein recruitment assay for demonstration and analysis of in vivo protein interactions in plant cells and its application to Tobacco mosaic virus movement protein.
abstract: We describe a simple fluorescent protein-based method to investigate interactions with a viral movement protein in living cells that relies on the in vivo re-localization of proteins in the presence of their interaction partners. We apply this method in combination with fluorescence lifetime imaging microscopy (FLIM) to demonstrate that a domain of the Tobacco mosaic virus (TMV) movement protein (MP) previously predicted to mediate protein:protein interactions is dispensable for these contacts. We suggest that this method can be generalized for analysis of other protein interactions in planta.
authors.affiliation: Institut de Biologie Moléculaire des Plantes du CNRS (UPR 2357), Université de Strasbourg, 12 rue du Général Zimmer, 67084 Strasbourg Cedex, France.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Boutant
authors.forename: Pascal
authors.initials: P
authors.lastname: Didier
authors.forename: Annette
authors.initials: A
authors.lastname: Niehl
authors.forename: Yves
authors.initials: Y
authors.lastname: Mély
authors.forename: Christophe
authors.initials: C
authors.lastname: Ritzenthaler
authors.forename: Manfred
authors.initials: M
authors.lastname: Heinlein
mesh_terms: D008164:Luminescent Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D053830:Plant Viral Movement Proteins / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000821:virology; D025941:Protein Interaction Mapping; D014026:Nicotiana / Q000235:genetics / Q000821:virology*; D014027:Tobacco Mosaic Virus / Q000378:metabolism*
chemical_list: D008164:Luminescent Proteins; D053830:Plant Viral Movement Proteins