Abstract
In eukaryotic cells, sphingoid long chain bases (LCBs) such as sphingosine or phytosphingosine (PHS) behave as second messengers involved in various processes including programmed cell death (PCD). In plants, induction of PCD by LCBs has now been described, but the signalling pathway is still enigmatic. Using Arabidopsis, we identify new key steps in this pathway. We demonstrate that PHS induces activation of the calcium-dependent kinase CPK3, which phosphorylates its binding partners, the 14-3-3 proteins. This phosphorylation leads to the disruption of the complex and to CPK3 degradation. Using cpk3 knockout lines, we demonstrate that CPK3 is a positive regulator of LCB-mediated PCD. These findings establish 14-3-3-regulated CPK3 as a key component of the LCB pathway leading to PCD in plants.
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Abbreviations
- CDPK or CPK:
-
calcium-dependent protein kinase
- DHS:
-
dihydrosphingosine
- DIG:
-
digoxigenin
- FB1:
-
fumonisin B1
- FDA:
-
fluorescein diacetate
- GAPDH:
-
glyceraldehyde-3-phosphate dehydrogenase
- LCB:
-
sphingoid long chain base
- MPK:
-
mitogen-activated protein kinase
- PCD:
-
programmed cell death
- PHS:
-
phytosphingosine
- PKA:
-
protein kinase A
- PKC:
-
protein kinase C
- SDK:
-
sphingosine-dependent protein kinase
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Acknowledgements
We thank Julian I Schroeder (University of California San Diego, CA, USA) for providing cpk3-1 seeds, Bernhard Wurzinger (University of Vienna, Austria) for providing cpk3-2 seeds, Ming-Che Shih (Academia Sinica, Taipei, Taiwan) for providing anti-GAPDH antibodies and Alexandre Perochon for providing CPK3 entry clone. We are grateful to Carol MacKintosh (MRC, Dundee, UK) for anti-14-3-3 antibodies and for other helpful tools. We thank Mercedes Pozuelo Rubio, Carole Pichereaux, Gisèle Borderies and Michel Rossignol for scientific discussions. We also thank Pauline Blanquet for help in experiments performed on plants. This work was supported by the Centre National de la Recherche Scientifique and the Université de Toulouse and was carried out in the Laboratoire de Recherche en Sciences Végétales, part of the Laboratoire d'Excellence entitled TULIP (ANR-10-LABX-41).
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Lachaud, C., Prigent, E., Thuleau, P. et al. 14-3-3-Regulated Ca2+-dependent protein kinase CPK3 is required for sphingolipid-induced cell death in Arabidopsis. Cell Death Differ 20, 209–217 (2013). https://doi.org/10.1038/cdd.2012.114
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DOI: https://doi.org/10.1038/cdd.2012.114
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