Protein phosphatase type 2C PP2CA together with ABI1 inhibits SnRK2.4 activity and regulates plant responses to salinity

[EN] Protein phosphatases 2C (PP2Cs) are important regulators of plant responses to abiotic stress. It is established that clade A PP2Cs inhibit ABA-activated SNF1-related protein kinases 2 (SnRK2s). Our recently published results show that ABI1, a member of clade A of PP2C is also a negative regulator of SnRK2.4, a kinase not activated in response to ABA. Here, we show that another member of this clade - PP2CA, interacts with and inhibits SnRK2.4. The salt-induced SnRK2.4/SnRK2.10 activity is higher in abi1-2 pp2ca-1 mutant than in wild type or single abi1 or pp2ca mutants, indicating that both phosphatases are inhibitors of SnRK2.4 and are at least partially redundant. Moreover, PP2CA together with ABI1 and SnRK2.4 regulates root growth in response to salinity.This work was supported by National Science Center (grant 2011/03/B/NZ3/00297 to GD). Funding in the laboratory of Pedro L. Rodriguez was provided by grant BIO2014-52537-R.Krzywinska, E.; Kulik, A.; Bucholc, M.; Antolín Fernández, M.; Rodríguez Egea, PL.; Dobrowolska, G. (2016). Protein phosphatase type 2C PP2CA together with ABI1 inhibits SnRK2.4 activity and regulates plant responses to salinity. Plant Signaling and Behavior. 11(12). https://doi.org/10.1080/15592324.2016.1253647S111

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

International audienceSeveral 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