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.
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.
Boutant
Emmanuel
E
Institut de Biologie Moléculaire des Plantes du CNRS (UPR 2357), Université de Strasbourg, 12 rue du Général Zimmer, 67084 Strasbourg Cedex, France.
Didier
Pascal
P
Niehl
Annette
A
Mély
Yves
Y
Ritzenthaler
Christophe
C
Heinlein
Manfred
M
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*
D008164:Luminescent Proteins; D053830:Plant Viral Movement Proteins