%0 Journal Article %1 liese2024imaging %A Liese, A. %A Eichstadt, B. %A Lederer, S. %A Schulz, P. %A Oehlschlager, J. %A Matschi, S. %A Feijo, J. A. %A Schulze, W. X. %A Konrad, K. R. %A Romeis, T. %D 2024 %J Plant Cell %K *Arabidopsis/metabolism myOwn %N 2 %P 276-297 %R 10.1093/plcell/koad196 %T Imaging of plant calcium-sensor kinase conformation monitors real time calcium-dependent decoding in planta %U https://www.ncbi.nlm.nih.gov/pubmed/37433056 %V 36 %X Changes in cytosolic calcium (Ca2+) concentration are among the earliest reactions to a multitude of stress cues. While a plethora of Ca2+-permeable channels may generate distinct Ca2+ signatures and contribute to response specificities, the mechanisms by which Ca2+ signatures are decoded are poorly understood. Here, we developed a genetically encoded Forster resonance energy transfer (FRET)-based reporter that visualizes the conformational changes in Ca2+-dependent protein kinases (CDPKs/CPKs). We focused on two CDPKs with distinct Ca2+-sensitivities, highly Ca2+-sensitive Arabidopsis (Arabidopsis thaliana) AtCPK21 and rather Ca2+-insensitive AtCPK23, to report conformational changes accompanying kinase activation. In tobacco (Nicotiana tabacum) pollen tubes, which naturally display coordinated spatial and temporal Ca2+ fluctuations, CPK21-FRET, but not CPK23-FRET, reported oscillatory emission ratio changes mirroring cytosolic Ca2+ changes, pointing to the isoform-specific Ca2+-sensitivity and reversibility of the conformational change. In Arabidopsis guard cells, CPK21-FRET-monitored conformational dynamics suggest that CPK21 serves as a decoder of signal-specific Ca2+ signatures in response to abscisic acid and the flagellin peptide flg22. Based on these data, CDPK-FRET is a powerful approach for tackling real-time live-cell Ca2+ decoding in a multitude of plant developmental and stress responses.

@article{liese2024imaging, abstract = {Changes in cytosolic calcium (Ca2+) concentration are among the earliest reactions to a multitude of stress cues. While a plethora of Ca2+-permeable channels may generate distinct Ca2+ signatures and contribute to response specificities, the mechanisms by which Ca2+ signatures are decoded are poorly understood. Here, we developed a genetically encoded Forster resonance energy transfer (FRET)-based reporter that visualizes the conformational changes in Ca2+-dependent protein kinases (CDPKs/CPKs). We focused on two CDPKs with distinct Ca2+-sensitivities, highly Ca2+-sensitive Arabidopsis (Arabidopsis thaliana) AtCPK21 and rather Ca2+-insensitive AtCPK23, to report conformational changes accompanying kinase activation. In tobacco (Nicotiana tabacum) pollen tubes, which naturally display coordinated spatial and temporal Ca2+ fluctuations, CPK21-FRET, but not CPK23-FRET, reported oscillatory emission ratio changes mirroring cytosolic Ca2+ changes, pointing to the isoform-specific Ca2+-sensitivity and reversibility of the conformational change. In Arabidopsis guard cells, CPK21-FRET-monitored conformational dynamics suggest that CPK21 serves as a decoder of signal-specific Ca2+ signatures in response to abscisic acid and the flagellin peptide flg22. Based on these data, CDPK-FRET is a powerful approach for tackling real-time live-cell Ca2+ decoding in a multitude of plant developmental and stress responses.}, added-at = {2024-02-15T15:08:22.000+0100}, author = {Liese, A. and Eichstadt, B. and Lederer, S. and Schulz, P. and Oehlschlager, J. and Matschi, S. and Feijo, J. A. and Schulze, W. X. and Konrad, K. R. and Romeis, T.}, biburl = {https://www.bibsonomy.org/bibtex/26db60c632067ee2fa2fc0365a039a2da/jvsi_all}, doi = {10.1093/plcell/koad196}, interhash = {be571d3527096bc4fb0044de69c4ad26}, intrahash = {6db60c632067ee2fa2fc0365a039a2da}, issn = {1532-298X (Electronic) 1040-4651 (Linking)}, journal = {Plant Cell}, keywords = {*Arabidopsis/metabolism myOwn}, note = {Liese, Anja Eichstadt, Bernadette Lederer, Sarah Schulz, Philipp Oehlschlager, Jan Matschi, Susanne Feijo, Jose A Schulze, Waltraud X Konrad, Kai R Romeis, Tina eng England 2023/07/11 Plant Cell. 2024 Jan 30;36(2):276-297. doi: 10.1093/plcell/koad196.}, number = 2, pages = {276-297}, timestamp = {2024-02-15T15:08:22.000+0100}, title = {Imaging of plant calcium-sensor kinase conformation monitors real time calcium-dependent decoding in planta}, type = {Journal Article}, url = {https://www.ncbi.nlm.nih.gov/pubmed/37433056}, volume = 36, year = 2024 }