Direct visualization of protein interactions in plant cells.
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.
Subramaniam
R
R
Department of Biochemistry, Université de Montréal, Montréal, Canada H3C 3J7.
Desveaux
D
D
Spickler
C
C
Michnick
S W
SW
Brisson
N
N
D000818:Animals; D001671:Biochemistry / Q000379:methods*; D002467:Cell Nucleus / Q000201:enzymology; D002469:Cell Separation; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000494:pharmacology; D007539:Isonicotinic Acids / Q000494:pharmacology; D051379:Mice; D008856:Microscopy, Fluorescence; D010944:Plants / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D013045:Species Specificity; D013053:Spectrophotometry; D013762:Tetrahydrofolate Dehydrogenase / Q000378:metabolism; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism
D005456:Fluorescent Dyes; D007539:Isonicotinic Acids; D011506:Proteins; D014157:Transcription Factors; C077261:2,6-dichloroisonicotinic acid; D013762:Tetrahydrofolate Dehydrogenase