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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
Author(s) -
Boutant Emmanuel,
Didier Pascal,
Niehl Annette,
Mély Yves,
Ritzenthaler Christophe,
Heinlein Manfred
Publication year - 2010
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2010.04126.x
Subject(s) - movement protein , tobacco mosaic virus , green fluorescent protein , fluorescence , protein–protein interaction , fluorescent protein , in vivo , biology , fluorescence microscope , microbiology and biotechnology , plant virus , bimolecular fluorescence complementation , biophysics , virus , coat protein , virology , biochemistry , genetics , gene , physics , rna , quantum mechanics
Summary 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.