The rice cold-responsive calcium-dependent protein kinase OsCPK17 is regulated by alternative splicing and post-translational modifications

M. Cecília Almadanim, Nuno M. Gonçalves, Margarida T.G. Rosa, Bruno M. Alexandre, André M. Cordeiro, Mafalda Rodrigues, Nelson J.M. Saibo, Cláudio M. Soares, Célia V. Romão, M. Margarida Oliveira, Isabel A. Abreu

Research output: Contribution to journalArticlepeer-review

22 Citations (Scopus)

Abstract

Plant calcium-dependent protein kinases (CDPKs) are key proteins implicated in calcium-mediated signaling pathways of a wide range of biological events in the organism. The action of each particular CDPK is strictly regulated by many mechanisms in order to ensure an accurate signal translation and the activation of the adequate response processes. In this work, we investigated the regulation of a CDPK involved in rice cold stress response, OsCPK17, to better understand its mode of action. We identified two new alternative splicing (AS) mRNA forms of OsCPK17 encoding truncated versions of the protein, missing the CDPK activation domain. We analyzed the expression patterns of all AS variants in rice tissues and examined their subcellular localization in onion epidermal cells. The results indicate that the AS of OsCPK17 putatively originates truncated forms of the protein with distinct functions, and different subcellular and tissue distributions. Additionally, we addressed the regulation of OsCPK17 by post-translational modifications in several in vitro experiments. Our analysis indicated that OsCPK17 activity depends on its structural rearrangement induced by calcium binding, and that the protein can be autophosphorylated. The identified phosphorylation sites mostly populate the OsCPK17 N-terminal domain. Exceptions are phosphosites T107 and S136 in the kinase domain and S558 in the C-terminal domain. These phosphosites seem conserved in CDPKs and may reflect a common regulatory mechanism for this protein family.

Original languageEnglish
Pages (from-to)231-246
Number of pages16
JournalBiochimica et Biophysica Acta - Molecular Cell Research
Volume1865
Issue number2
DOIs
Publication statusPublished - 1 Feb 2018

Keywords

  • Alternative splicing
  • CDPK
  • Phosphorylation
  • Post-translational modifications
  • Rice
  • Subcellular localization

Fingerprint

Dive into the research topics of 'The rice cold-responsive calcium-dependent protein kinase OsCPK17 is regulated by alternative splicing and post-translational modifications'. Together they form a unique fingerprint.

Cite this