Plant Molecular Biology

, Volume 63, Issue 4, pp 491–503 | Cite as

Different phosphorylation mechanisms are involved in the activation of sucrose non-fermenting 1 related protein kinases 2 by osmotic stresses and abscisic acid

  • Marie Boudsocq
  • Marie-Jo Droillard
  • Hélène Barbier-Brygoo
  • Christiane LaurièreEmail author


In Arabidopsis cell suspension, hyperosmotic stresses (mannitol and NaCl) were previously shown to activate nine sucrose non-fermenting 1 related protein kinases 2 (SnRK2s) whereas only five of them were also activated by abscisic acid (ABA) treatment. Here, the possible activation by phosphorylation/dephosphorylation of each kinase was investigated by studying their phosphorylation state after osmotic stress, using the Pro-Q Diamond, a specific dye for phosphoproteins. All the activated kinases were phosphorylated after osmotic stress but the induced phosphorylation changes were clearly different depending on the kinase. In addition, the increase of the global phosphorylation level induced by ABA application was lower, suggesting that different mechanisms may be involved in SnRK2 activation by hyperosmolarity and ABA. On the other hand, SnRK2 kinases remain activated by hyperosmotic stress in ABA-deficient and ABA-insensitive mutants, indicating that SnRK2 osmotic activation is independent of ABA. Moreover, using a mutant form of SnRK2s, a specific serine in the activation loop was shown to be phosphorylated after stress treatments and essential for activity and/or activation. Finally, SnRK2 activity was sensitive to staurosporine, whereas SnRK2 activation by hyperosmolarity or ABA was not, indicating that SnRK2 activation by phosphorylation is mediated by an upstream staurosporine-insensitive kinase, in both signalling pathways. All together, these results indicate that different phosphorylation mechanisms and at least three signalling pathways are involved in the activation of SnRK2 proteins in response to osmotic stress and ABA.


Abscisic acid Arabidopsis thaliana Osmotic stresses Phosphorylation Pro-Q® Diamond SnRK2 



abscisic acid


myelin basic protein


sucrose non-fermenting 1 related protein kinase 2



We thank Dr. Pascale Bertrand and Dr. Yannick Saintigny for kind introduction to the Typhoon imaging system. We also thank Dr. Sylvain Merlot and Dr. Helen North for kindly providing abi1-1 and aba1-3 seeds, respectively.


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Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • Marie Boudsocq
    • 1
    • 2
    • 3
  • Marie-Jo Droillard
    • 1
  • Hélène Barbier-Brygoo
    • 1
  • Christiane Laurière
    • 1
    Email author
  1. 1.Institut des Sciences du Végétal, UPR 2355, CNRSGif sur Yvette CedexFrance
  2. 2.Department of GeneticsHarvard Medical SchoolBostonUSA
  3. 3.Department of Molecular BiologyMassachusetts General HospitalBostonUSA

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