Rice calcium-dependent protein kinase OsCPK17 targets plasma membrane intrinsic protein and sucrose phosphate synthase and is required for a proper cold stress response
AuthorsAlmadanim, M. Cecília
Alexandre, Bruno M.
Rosa, Margarida T.G.
Leitão, António E.
Ramalho, José C.
Lam, TuKiet T.
Abreu, Isabel A.
Oliveira, M. Margarida
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AbstractCalcium-dependent protein kinases (CDPKs) are involved in plant tolerance mechanisms to abiotic stresses. Although CDPKs are recognized as key messengers in signal transduction, the specific role of most members of this family remains unknown. Here we test the hypothesis that OsCPK17 plays a role in rice cold stress response by analyzing OsCPK17 knockout, silencing, and overexpressing rice lines under low temperature. Altered OsCPK17 gene expression compromises cold tolerance performance, without affecting the expression of key cold stress-inducible genes. A comparative phosphoproteomic approach led to the identification of six potential in vivo OsCPK17 targets, which are associated with sugar and nitrogen metabolism, and with osmotic regulation. To test direct interaction, in vitro kinase assays were performed, showing that the sucrose phosphate synthase OsSPS4, and the aquaporin OsPIP2;1/OsPIP2;6 are phosphorylated by OsCPK17 in a calcium-dependent manner. Altogether, our data indicates that OsCPK17 is required for a proper cold stress response in rice, likely affecting the activity of membrane channels and sugar metabolism.
CitationAlmadanim MC, Alexandre BM, Rosa MTG, Sapeta H, Leitão AE, et al. (2017) Rice calcium-dependent protein kinase OsCPK17 targets plasma membrane intrinsic protein and sucrose phosphate synthase and is required for a proper cold stress response. Plant, Cell & Environment. Available: http://dx.doi.org/10.1111/pce.12916.
SponsorsWe acknowledge: Alexandra Marques for cloning OsCPK17 into pDEST28; Pedro Pereira for the help with the transgenic rice plants; Nuno Gonçalves, Mafalda Rodrigues and Vanessa Azevedo for the aid with electrolyte leakage datapoint sample collection; Tiago Lourenço and Ana Paula Farinha for providing some of the primers used; Margarida Saramago for the support with scintillation counter equipment and borrowed material; Edward Voss, Jean Kanyo, and Kathrin Wilczak for their assistance in MS sample preparation, data collection, and initial data analysis, respectively. We also thank Rossana Henriques (CRAG), Nelson Saibo (ITQB), and Alison Funston (Monash University, Australia) for critical revision. This research was funded by Portuguese Fundação para a Ciência e a Tecnologia: grants SFRH/BD/61121/2009 to M.C.A., SFRH/BD/84219/2012 to M.T.G.R., SFRH/BD/89781/2012 to H.S., SFRH/BPD/98619/2013 to B.M.A.; FCT Investigator (POPH-QREN) to I.A.A.. The work was also supported by the FCT research units GREEN-it
JournalPlant, Cell & Environment