Regulation of Nicotiana tabacum osmotic stress-activated protein kinase and its cellular partner GAPDH by nitric oxide in response to salinity

Izabela Wawer , Maria Bucholc , Jéremy Astier , Anna Anielska-Mazur , Jennifer Dahan , Anna Kulik , Aleksandra Wysłouch-Cieszynska , Monika Zaręba-Kozioł , Ewa Krzywinska , Michal Dadlez , Grażyna Dobrowolska , David Wendehenne (1)

1 PMEBBCE - Plante - microbe - environnement : biochimie, biologie cellulaire et écologie

Several studies focused on elucidating the mechanism of nitric oxide (NO) signalling in plant cells highlighted that its biological effects are partly mediated by protein kinases. Identity of these kinases and details of how NO modulates their activities, however, remain poorly investigated. Here, we have attempted to clarify the mechanisms underlying NO action in regulation of Nicotiana tabacum osmotic stress-activated protein kinase (NtOSAK), a member of the SNF1-related protein kinase 2 family. We found that in tobacco BY-2 cells exposed to salt stress, NtOSAK is rapidly activated partly through a NO-dependent process. This activation, as well as the one observed following treatment of BY-2 cells with the NO donor DEA/NO, involved the phosphorylation of two residues located in the kinase activation loop, one being identified as Ser-158. Our results indicate that NtOSAK does not undergo the direct chemical modifications of its Cys residues by S-nitrosylation. Using a co-immunoprecipitation-based strategy, we identified several proteins present in immunocomplex with NtOSAK in salt-treated cells including the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Our results indicate that NtOSAK directly interacts with GAPDH in planta. Furthermore, in response to salt, GAPDH showed a transient increase of its S-nitrosylation level which was correlated with the time course of NtOSAK activation. However, GADPH S-nitrosylation did not influence its interaction with NtOSAK and did not impact the activity of the protein kinase. Taken together, the results support the hypothesis that NtOSAK and GAPDH form a cellular complex and that both proteins are regulated directly or indirectly by NO.

Mots clés: Life Sciences

Domaines: Biochimie, Biologie Moléculaire

Izabela Wawer, Maria Bucholc, Jéremy Astier, Anna Anielska-Mazur, Jennifer Dahan, et al.. Regulation of Nicotiana tabacum osmotic stress-activated protein kinase and its cellular partner GAPDH by nitric oxide in response to salinity. Biochemical Journal, 2010, 429 (1), pp.73-83. ⟨10.1042/BJ20100492⟩. ⟨hal-00491634⟩