Functional study of phosphorylation-dependent interaction between 14-3-3 and glutamate receptor-like gene 3.7 in Arabidopsis thaliana
Date Issued
2013
Date
2013
Author(s)
Wang, Po-Hsun
Abstract
Ionotropic glutamate receptors (iGluRs) are ligand-gated cation channels that mediate fast excitatory neurotransmission in the mammalian central nervous system. In the model plant Arabidopsis thaliana, a family of 20 proteins share similarities to animal iGluRs in sequence and predicted secondary structure. AtGLRs function as non-selective cation channel to transport cations. Previous researches showed biological functions of glutamate receptor in root elongation and abotic stress response. However, the signal transduction pathway in plants is still not clear. 14-3-3 protein is a scaffold protein, which can interact with ion channel in the cytosol and change their ion permeability. 14-3-3 protein binds to phospho-serine/threonine amino acid in a sequence-specific manner. It has been reported that 14-3-3 protein may interact with GLR3.7. Besides, calcium-dependent protein kinase (CDPK) can phosphorylate GLR3.7 in virto. To confirm whether GLR3.7 phosphorylation site overlapping with the 14-3-3 binding site, we used in vitro kinase assay to identify the phosphorylation site in GLR3.7 phosphorylated by CDPK. Then we observed the subcellular localization of AtGLR3.7 using confocol microscope. Then we performed bimolecular fluorescence complementation (BiFC) assay, pull-down, quartz crystal microbalance analysis and co-immunoprecipitation to confirm 14-3-3 binding site. In addition, we found that AtGLR3.7 is involved in root hair development, and the results revealed that AtGLR3.7 may regulate root hair elongation. Overall, these findings demonstrate that CDPK phosphorylates Ser-860 of AtGLR3.7, and the phosphorylation site can be bound by 14-3-3 protein to activate the channel activity and influence root hair development in Arabidopsis thaliana.
Subjects
阿拉伯芥
GLR3.7
陽離子通道
鈣離子依存性蛋白激酶
磷酸化
14-3-3
根毛
Type
thesis
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