Influence of sucrose starvation, osmotic and salt stresses on expression profiles of genes involved in the development of autophagy by means of microtubules
Abstract
Aim. The aim of this work was to investigate changes in expression profiles of key genes involved in the development of autophagy by means of microtubules under the influence of sucrose starvation, osmotic and salt stresses. Methods. Arabidopsis thaliana seeds were sown aseptically on Murashige and Skoog solid medium. Salt and osmotic stresses were simulated by seed germination and seedlings cultivation on the media containing 150 mМ NaCl and 10 mМ mannitol, respectively. For investigation of starvation-induced autophagy plants were germinated and grown on sucrose-free medium. Results. Changes in expression of α-tubulin and atg8 genes had clearly defined stressdependent nature. Overexpression of tua1 and atg8e under starvation; tua3 and atg8f under osmotic stress; tua3 and atg8f, atg8e during salt stress indirectly testifies interaction between the structural units of autophagosomes and microtubules. It was shown that influence of investigated abiotic stimuli results in overexpression of elp3 and hda6 genes. Small increase in expression levels of hexokinase 2 and 3 was demonstrated. Conclusions. Transcriptome analysis of key genes involved in realization of autophagy induced by sucrose starvation, osmotic and salt stresses in Arabidopsis thaliana cells was conducted. Received data indirectly testifies interaction between the structural units of autophagosomes and microtubules and enables to point α-tubulin and atg8 genes, which are specific for the realization of autophagy induced by a certain abiotic stimuli. Expression profiles of elp3/deacetylases as well as hexokinases indicate the critical role of α-tubulin acetylation for autophagic response, that is involved in the development of programmed cell death.
Keywords: autophagy, sucrose starvation, osmotic stress, salt stress, transcriptome analysis, α-tubulin, atg8.
References
Mahajan S., Tuteja N. Cold, salinity and drought stresses: an overview. Arch. Biochem. Biophys. 2005. Vol. 444. P.139-158. doi: 10.1016/j.abb.2005.10.018
Zhu J.K. Cell signaling under salt, water and cold stresses. Curr. Opin. Plant Biol. 2001. Vol. 4. P.401-406.
Batelli G., Verslues P.E., Agius F., Qiu Q., Fujii H., Pan S., et al. SOS2 promotes salt tolerance in part by interacting with the vacuolar H+-ATPase and upregulating its transport activity. Mol. Cell Biol. 2007. Vol. 27. P.7781-7790. doi: 10.1128/MCB.00430-07
Moriyasu Y., Ohsumi Y. Autophagy in tobacco suspension-cultured cells in response to sucrose starvation. Plant Physiol. 1996. Vol. 111. P. 1233-1241.
Slavikova S., Ufaz S., Avin-Wittenberg T., Levanony H., Galili G. An autophagy-associated Atg8 protein is involved in the responses of Arabidopsis seedlings to hormonal controlsand abiotic stresses. J. Exp. Bot. 2008. Vol. 59. P. 4029-4043. doi: 10.1093/jxb/ern244
Liu Y., Xiong Y., Bassham D.C. Autophagy is required for tolerance of drought and salt stress in plants. Autophagy. 2009. Vol. 5(7). P.954-963.
Monetti E., Kadono T., Tran D., Azzarello E., Arbelet-Bonnin D., Biligui B., Briand J., Kawano T., Mancuso S., Bouteau F. Deciphering early events involved in hyperosmotic stress-induced programmed cell death in tobacco BY-2 cells. J Exp Bot. 2014. Vol. 65(5). P. 1361-1375. doi: 10.1093/jxb/ert460
Lin J., Wang Y., Wang G. Salt stress-induced programmed cell death in tobacco protoplasts is mediated by reactive oxygen species and mitochondrial permeability transition pore status. J. Plant Physiol. 2006. Vol. 163(7). P. 731-739. doi: 10.1016/j.jplph.2005.06.016
Fedyna V., Lytvyn D., Yemets A., Blume Ya. Microtubule cytoskeleton participates in realization of autophagy, which is crucial for cell surviving under the influence of stressful stimuli in Arabidopsis thaliana. Factors Exp. Evol. Organisms. 2016. Vol. 19. P.47-50 (in Ukr.).
Wang C., Li J., Yuan M. Salt tolerance requires cortical microtubule reorganization in Arabidopsis. Plant Cell Physiol. 2007. Vol. 48(11). P. 1534-1547. doi: 10.1093/pcp/pcm123
Blancaflor E.B., Hasenstein K.H. Growth and microtubule orientation of Zea mays roots subjected to osmotic stress. Int. J. Plant Sci. 1995. Vol. 156(6). P. 774-783.
Geeraert C., Ratier A., Pfisterer S.G., et al. Starvation-induced hyperacetylation of tubulin is required for the stimulation of autophagy by nutrient deprivation. J. Biol. Chem. 2012. Vol. 285(31). P. 24184-24194. doi: 10.1074/jbc.M109.091553
Lytvyn D., Yemets A., Bergounioux C., Blume Ya. B. Development of autophagy in BY-2 (Nicotiana tabacum) cells is accompanied by acetylation of α-tubulin. Reports Natl. Acad. Sci. Ukraine. 2013. No 2. P. 179-185 (in Ukr).
Lytvyn D.I., Blume Ya.B. Microtubular cytoskeleton in autophagy and programmed cell death development in plants. In: Programmed Cell Death in Plants and Animals (Ed. J. Rice), 2016, Nova Science Publishers: New York, p. 1-26.
Olenieva V., Lytvyn D., Yemets A., Bergounioux C., Blume Y. Tubulin acetylation accompanies autophagy development induced by different abiotic stimuli in Arabidopsis thaliana. Cell Biol. Int. 2017. doi: 10.1002/cbin.10842
Tran H.T., Nimick M., Uhrig R.G., Templeton G., Morrice N., Gourlay R., DeLong A., Moorhead G.B. Arabidopsis thaliana histone deacetylase 14 (HDA14) is an α-tubulin deacetylase that associates with PP2A and enriches in the microtubule fraction with the putative histone acetyltransferase ELP3. Plant J. 2012. Vol. 71(2). P. 263-272. doi: 10.1111/j.1365-313X.2012.04984.x
Olenieva V., Lytvyn D., Yemets A., Blume Ya. B. Influence of UV-B on expression profiles of genes involved in the development of autophagy by means of microtubules. Reports Natl. Acad. Sci. Ukraine. 2018, in press (in Ukr).
Lee J.Y., Koga H., Kawaguchi Y., et al. HDAC6 controls autophagosome maturation essential for ubiquitin-selective quality-control autophagy. EMBO J. 2010. Vol. 29(5). P. 969-980. doi: 10.1038/emboj.2009.405
