In vivo identification of putative CPK5 substrates in Arabidopsis thaliana
| Autor: | Nicolas Valentin, Jean Colcombet, Heribert Hirt, Liliana Avila-Ospina, Marlène Davanture, Marie Boudsocq, Naganand Rayapuram, Tiffany Yip Delormel, Julien Lang, Michel Zivy |
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| Přispěvatelé: | Institut des Sciences des Plantes de Paris-Saclay (IPS2 (UMR_9213 / UMR_1403)), Université d'Évry-Val-d'Essonne (UEVE)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Génétique Quantitative et Evolution - Le Moulon (Génétique Végétale) (GQE-Le Moulon), AgroParisTech-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), King Abdullah University of Science and Technology (KAUST), Université d'Évry-Val-d'Essonne (UEVE)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) |
| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
Arabidopsis Plant Science Biology Genes Plant 01 natural sciences 03 medical and health sciences Gene Expression Regulation Plant Genetics AtCPK5 Arabidopsis thaliana [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology substrates 030304 developmental biology Calcium signaling 0303 health sciences calcium Abiotic stress Kinase phosphorylation General Medicine [SDV.BV.BOT]Life Sciences [q-bio]/Vegetal Biology/Botanics biology.organism_classification Cell biology Ubiquitin ligase Cytosol biology.protein Phosphorylation Agronomy and Crop Science Protein Kinases 010606 plant biology & botany |
| Zdroj: | Plant Science Plant Science, 2022, 314, pp.111121. ⟨10.1016/j.plantsci.2021.111121⟩ Plant Science, Elsevier, 2022, 314, pp.111121. ⟨10.1016/j.plantsci.2021.111121⟩ |
| ISSN: | 1873-2259 0168-9452 |
| Popis: | International audience; Calcium signaling mediates most developmental processes and stress responses in plants. Among plant calcium sensors, the calcium-dependent protein kinases display a unique structure harboring both calcium sensing and kinase responding activities. AtCPK5 is an essential member of this family in Arabidopsis that regulates immunity and abiotic stress tolerance. To understand the underlying molecular mechanisms, we implemented a biochemical approach to identify in vivo substrates of AtCPK5. We generated transgenic lines expressing a constitutively active form of AtCPK5 under the control of a dexamethasone-inducible promoter. Lines expressing a kinase-dead version were used as a negative control. By comparing the phosphoproteome of the kinase-active and kinase-dead lines upon dexamethasone treatment, we identified 5 phosphopeptides whose abundance increased specifically in the kinase-active lines. Importantly, we showed that all 5 proteins were phosphorylated in vitro by AtCPK5 in a calcium-dependent manner, suggesting that they are direct targets of AtCPK5. We also detected several interaction patterns between the kinase and the candidates in the cytosol, membranes or nucleus, consistent with the ubiquitous localization of AtCPK5. Finally, we further validated the two phosphosites S245 and S280 targeted by AtCPK5 in the E3 ubiquitin ligase ATL31. Altogether, those results open new perspectives to decipher AtCPK5 biological functions. |
| Databáze: | OpenAIRE |
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