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Direct visualization of protein interactions in plant cells

Abstract

The protein NPR1/NIM1 is required for the induction of systemic acquired resistance (SAR) in plants and has been shown to interact with members of the TGA/OBF family of basic leucine zipper (bZIP) transcription factors. However, to date, there is no method available to monitor such interactions in plant cells. We report here an in vivo protein fragment complementation assay (PCA), based on association of reconstituted murine dihydrofolate reductase (mDHFR) with a fluorescent probe to detect protein–protein interaction in planta. We demonstrate that the interaction between Arabidopsis NPR1/NIM1 and the bZIP factor TGA2 is induced by the regulators of SAR, salicylic acid (SA) and its analog 2,6-dichloroisonicotinic acid (INA) with distinct species-specific responses. Furthermore, the induced interaction is localized predominantly in the nucleus. Protein fragment complementation assays could be of value to agricultural research by providing a system for high-throughput biochemical pathway mapping and for screening of small molecules that modulate protein interactions.

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Acknowledgements

We are very grateful to Dr. Pierre Fobert, National Research Council of Canada, Saskatoon, for providing Arabidopsis NPR1/NIM1 and TGA2 and Dr. Charles Després for useful discussions. We thank Dr. Daniel Matton for the plasmid containing the double 35S enhancer element of CaMV. D.D. was the recipient of a scholarship from the Natural Sciences and Engineering Research Council of Canada (NSERC). This work was supported by grants from NSERC to S.W.M. and N.B. and from the Fonds Concerté d'Aide à la Recherche du Québec to N.B.

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Subramaniam, R., Desveaux, D., Spickler, C. et al. Direct visualization of protein interactions in plant cells. Nat Biotechnol 19, 769–772 (2001). https://doi.org/10.1038/90831

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