Structure and function of NtCDPK1, a calcium-dependent protein kinase in tobacco

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Structure and function of NtCDPK1, a calcium-dependent protein kinase in tobacco
Gyeong Mee Yoon; Sang Sook Lee; Hyun Sook Pai
Bibliographic Citation
Journal of Plant Biotechnology, vol. 2, no. 2, pp. 79-82
Publication Year
We have isolated a cDNA encoding' a calcium-dependent protein kinase (CDPK) in Nicotiana tabacum, which was designated NtCDPKl. Accumulation of the NtCDPKl mRNA was stimulated by various stimuli, including phytohormones, CaCl2J wounding, fungal elicitors, chitin and methyl jasmonate. The NtCDPKl gene encodes a functional Serffhr protein kinase of which phosphoηlation activity is strongly induced by calcium. By analyzing expression of the NtCDPK1-GFP fusion protein and by immunoblotting with antibody which reacts with NtCDPK1, we found that NtCDPK1 is localized in membrane and nucleus in plant cells. Silencing expression of the NtCDPKl transgene resulted in marked decrease of lateral root development in the transgenic tobacco plants. Yeast two hybrid screening using NtCDPK1 as a bait identified a tobacco homologue of proteasome regulatory subunit 21D7, designated Nt21D7. The 21D7 mRNA has been shown to be predominantly expressed in proliferating tissues in the cell cycledependent manner in carrot. The recombinant NtCDPK1 protein associated with Nt21D7 in vitro, and could phosphorylate the Nt21D7 protein in vitro in the presence of calcium, suggesting that Nt 21D7 protein is a natural substrate of NtCDPK1 in tobacco. These results suggest that NtCDPK1 may regulate cell proliferation processes, such as lateral root formation, by regulating specificity and/or activity of proteasome-mediated protein degradation pathway.
Korea Soc-Assoc-Inst
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