title: Rice calcium-dependent protein kinase OsCPK17 targets plasma membrane intrinsic protein and sucrose-phosphate synthase and is required for a proper cold stress response.
abstract: Calcium-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 analysing 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.
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, 2780-157, Oeiras, Portugal.
authors.forename: M Cecília
authors.initials: MC
authors.lastname: Almadanim
authors.affiliation: Instituto de Biologia Experimental e Tecnológica, 2780-157, Oeiras, Portugal.
authors.forename: Bruno M
authors.initials: BM
authors.lastname: Alexandre
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, 2780-157, Oeiras, Portugal.
authors.forename: Margarida T G
authors.initials: MTG
authors.lastname: Rosa
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, 2780-157, Oeiras, Portugal.
authors.forename: Helena
authors.initials: H
authors.lastname: Sapeta
authors.affiliation: Plant Stress and Biodiversity, Linking Landscape, Environment, Agriculture and Food (LEAF), Dept. Recursos Naturais, Ambiente e Território (DRAT), Instituto Superior de Agronomia, Universidade de Lisboa, 2784-505, Oeiras, Portugal.
authors.forename: António E
authors.initials: AE
authors.lastname: Leitão
authors.affiliation: Plant Stress and Biodiversity, Linking Landscape, Environment, Agriculture and Food (LEAF), Dept. Recursos Naturais, Ambiente e Território (DRAT), Instituto Superior de Agronomia, Universidade de Lisboa, 2784-505, Oeiras, Portugal.
authors.forename: José C
authors.initials: JC
authors.lastname: Ramalho
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, 06520-8024, USA.|MS and Proteomics Resource, WM Keck Foundation Biotechnology Resource Laboratory, Yale University, New Haven, CT, 06520-8024, USA.
authors.forename: TuKiet T
authors.initials: TT
authors.lastname: Lam
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, 2780-157, Oeiras, Portugal.|Division of Biological and Environmental Sciences and Engineering, King Abdullah University of Science and Technology (KAUST), Thuwal, 23955-6900, Saudi Arabia.
authors.forename: Sónia
authors.initials: S
authors.lastname: Negrão
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, 2780-157, Oeiras, Portugal.|Instituto de Biologia Experimental e Tecnológica, 2780-157, Oeiras, Portugal.
authors.forename: Isabel A
authors.identifier: http://orcid.org/0000-0002-5566-2146
authors.initials: IA
authors.lastname: Abreu
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, 2780-157, Oeiras, Portugal.|Instituto de Biologia Experimental e Tecnológica, 2780-157, Oeiras, Portugal.
authors.forename: M Margarida
authors.initials: MM
authors.lastname: Oliveira
mesh_terms: D020346:Aquaporins / Q000235:genetics / Q000378:metabolism; D058639:Cold-Shock Response / Q000502:physiology*; D018506:Gene Expression Regulation, Plant; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism*; D012275:Oryza / Q000502:physiology*; D010766:Phosphorylation; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D011494:Protein Kinases / Q000378:metabolism*; D012639:Seeds / Q000254:growth & development / Q000378:metabolism; D017871:Calcium-Calmodulin-Dependent Protein Kinases
chemical_list: D020346:Aquaporins; D005964:Glucosyltransferases; D010940:Plant Proteins; D011494:Protein Kinases; C022698:sucrose-phosphate synthase; C454865:calcium-dependent protein kinase, rice; D017871:Calcium-Calmodulin-Dependent Protein Kinases
keywords: CDPK; abiotic stress; phosphoproteomics; phosphorylation; signal transduction