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bioRxiv · 10.1101/2023.03.13.532409

Imaging of plant calcium-sensor kinase conformation monitors real time calcium decoding in planta

Liese, A.·Eichstaedt, B.·Lederer, S.·Schulz, P.·Oehlschlaeger, J.·Feijo, J. A.·Schulze, W. X.·Konrad, K. R.·Romeis, T.
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Abstract

Changes in cytosolic calcium concentration are among the earliest reactions to a multitude of stress cues. Whereas a plethora of calcium-permeable channels may generate distinct calcium signatures and contribute to response specificities, the mechanisms by which calcium signatures are decoded is poorly understood. Here we develop a genetically encoded, FRET-based reporter that visualizes the conformational change of calcium-dependent protein kinases (CDPKs/CPKs), preceding kinase activation, for calcium-dependent AtCPK21 and calcium-independent AtCPK23. In pollen tubes, naturally displaying a physiological calcium range, CPK21-FRET, but not CPK23-FRET, report activity oscillations with similar features to cytosolic calcium, suggesting an isoform-specific calcium dependency and reversibility of the conformational change. In guard cells CPK21-FRET identifies CPK21 as a decoder of signal-specific calcium signatures in response to ABA and flg22. Based on this data, CDPK-FRET stands as a novel approach for tackling real-time live-cell calcium decoding in a multitude of plant developmental and stress responses.

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Liese, A., Eichstaedt, B., Lederer, S., Schulz, P., Oehlschlaeger, J., Feijo, J. A., Schulze, W. X., Konrad, K. R., Romeis, T.. 2023-03-13. Imaging of plant calcium-sensor kinase conformation monitors real time calcium decoding in planta. https://doi.org/10.1101/2023.03.13.532409

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