Understanding the molecular mechanisms involved in cell cycle control is essential for developmental studies, specially in plants , where most morphogenesis takes place after embryogenesis. Protein kinase CK2 is part of the complex network of protein kinases that control cell division. CK2 activity increases rapidly after stimulation of cell division, and it shows cell-cycle dependent regulation, with peaks of activity in G1/S and M. Results in our laboratory show that CK2 activity is regulated by transcriptional, post-trasncriptional and post-transductional mechanisms. To get a deeper understanding of the role of CK2 in plant's cell division we propose the following objectives: 1) Post-transductional control of CK2 activity: subcellular localization of the enzyme during the cell cycle in BY2 cells, functional role of the N-terminal extension of CK2b, and role of the proteosome in the control of CK2b levels. 2) Inhibition of CK2 activity by overexpression of a dominant negative mutant of CK2a using an inducible promoter. According to our previous results, the use of this mutant will allow us to explore the role of CK2 in G1/S and to identify substrates specific of this particular phase of the cell cycle. Introduction of this construct in Arabidopsis plants will allow us to study the relationship between CK2 activity depletion and development. Our group has been also working for years with the enzyme glutathione-dependent formaldehyde dehydrogenase. Transgenic Arabidopsis plants that overexpress the enzyme, generated in our laboratoy, show a very significant inhibition of root growth that might be due to depletion of GSH. An additional goal of this project is to study this interesting phenotype, that might establish a link between cell division and oxidative stress.
|Effective start/end date||13/12/04 → 13/12/07|
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