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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 |
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| Authors: | Emmanuel Boutant Pascal Didier Annette Niehl Yves Mély Christophe Ritzenthaler Manfred Heinlein |
| Affiliation: | 1. Institut de Biologie Moléculaire des Plantes du CNRS (UPR 2357), Université de Strasbourg, 12 rue du Général Zimmer, 67084 Strasbourg Cedex, France;2. Present address: EMBL, Meyerhofstrasse 1, 69117 Heidelberg, Germany.;3. Laboratoire Biophotonique et Pharmacologie, UMR CNRS 7213, Faculté de Pharmacie, Université de Strasbourg, 74 route du Rhin, 67401 Illkirch, France;4. Department of Plant Physiology, Botanical Institute, University of Basel, Hebelstrasse 1, 4056 Basel, Switzerland |
| 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. |
| Keywords: | Tobacco mosaic virus movement protein in vivo protein interactions GFP |