SUMMARY. SnRK1 protein kinases are an integral component of cellular signaling. These kinases are also involved in formation of stress response, energy metabolism regulation, seed germination and maturation, autophagy, etc. However, many of these protein kinases functions remain unknown. In order to study the intracellular localization of KIN10 (the catalytic subunit of the protein kinase complexes of SnRK1), transformation of the wild ecotype of A. thaliana (Col-0) protoplasts by genetic construct containing the KIN10-RFP fusion gene was performed. It was detected that the fusion protein KIN10-RFP is diffusely distributed in the cytoplasm, however, it is mainly localized on the periphery, closely to plasma membrane. Using immunofluorescence microscopy, diffuse localization of KIN10 and γ-tubulin in the cytoplasm of A. thaliana root cells was demonstrated. Differences in the intracellular localization of γ-tubulin in the root cells of the kin10, kin11 and wild-type A. thaliana knockout roots were found. In both mutants, a low fluorescence intensity of γ-tubulin was observed compared it comparison with wild type plants. These patterns of intracellular distribution may indicate certain interruption in the process of their γ-tubulin complexes formation. The low fluorescence intensity of γ-tubulin was also detected in the cells of all studied lines grown under energy deficit conditions. In particular, the lowest level of fluorescence was recorded in the kin10 and kin11 mutants under the these stress conditions. It may indicate the influence of the synergistic effect of simultaneous dysfunction of one of these genes and energy deficiency on γ-tubulin complexes (γTuSC and γTuRC) formation. The obtained results indicate the possible involvement of SnRK1 (KIN10 and KIN11) in the physiological response of plants to energy stress. Considering our bioinformatics analysis regarding the possible phosphorylation of plant γ-tubulin KIN10 for the Ser131 residue, it can be assumed that the protein kinase SnRK1 involved in the regulation of microtubule polymerization in plants.
Keywords: SnRK1, protein kinases, γ-tubulin, cell localization, KIN10, KIN11, mutant lines
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