14-3-3蛋白與中鹼性蔗糖轉化酶在阿拉伯芥中分子間之交互作用

Physical interaction between 14-3-3 and neutral/alkaline invertase in Arabidopsis thaliana

賴軒正(Shiuan-Jeng Lai)

指導教授 ： 張英峯

國立臺灣大學/生命科學院/植物科學研究所/碩士(2011年)

https://doi.org/10.6342/NTU.2011.01512

摘要

14-3-3蛋白是一種鷹架蛋白，在阿拉伯芥中可以調控許多蛋白質的功能，例如酵素的活性，還有影響蛋白質在細胞內的分布。在前人的研究中，藉由雙重性親合純化發現了14-3-3蛋白跟中性蔗糖轉化酶有交互作用。蔗糖轉化酶是一種可以水解蔗糖轉化為葡萄糖與果糖的酵素，我們認為中性蔗糖轉化酶可能是間接的或是直接的與14-3-3有交互作用，因此藉由雙分子螢光互補分析確定中性蔗糖轉化酶與14-3-3有直接的交互作用，並且藉由T-DNA 插入變異株來研究中性蔗糖轉化酶可能的功能。結果證實了14-3-3與中性蔗糖轉化酶會在植物細胞中有直接的交互作用，進而可能影響蛋白質在胞器間的穿梭，而中性蔗糖轉化酶可能對於抵抗鹽分逆境有幫助。

關鍵字

中性蔗糖轉化酶14-3-3 ； 鹽分逆境 ； 蛋白質磷酸化 ； 雙分子螢光互補系統

並列摘要

14-3-3 is a scaffold protein which regulates many protein functions such as enzyme activity and protein localization in Arabidopsis. In a previous study, 14-3-3 was found to interact with neutral invertase CINV1 by use of tandem affinity purification as a tool. We hypothesized that neutral invertase CINV1 may directly or indirectly interact with 14-3-3. Invertase catalyzes sucrose catabolism into glucose and fructose. We test and confirmed the interaction between 14-3-3omega and neutral invertase CINV1 by Bimolecular Fluorescence Complementation. This interaction may change the subcellular localization of CINV1. We have examined the phenotype of Arabidopsis cinv1 mutants under various growth condition. Our results indicate that the cinv1 mutants are salt sensitive.

並列關鍵字

14-3-3 ； cytosolic invertase ； Bimolecular Fluorescence Complementation ； salt stress

參考文獻

Aitken, A. (2006). 14-3-3 proteins: A historic overview. Semin Cancer Biol 16: 162-172.

Arnold, T.M., and Schultz, J.C. (2002). Induced sink strength as a prerequisite for induced tannin biosynthesis in developing leaves of Populus. Oecologia 130: 585-593.

Balibrea, M.E., Cuartero, J., Bolarin, M.C., and Perez-Alfocea, F. (2003). Sucrolytic activities during fruit development of Lycopersicon genotypes differing in tolerance to salinity. Physiol Plant 118: 38-46.

Barratt, D.H., Derbyshire, P., Findlay, K., Pike, M., Wellner, N., Lunn, J., Feil, R., Simpson, C., Maule, A.J., and Smith, A.M. (2009). Normal growth of Arabidopsis requires cytosolic invertase but not sucrose synthase. Proc Natl Acad Sci U S A 106: 13124-13129.

Baumert, A., Mock, H.P., Schmidt, J., Herbers, K., Sonnewald, U., and Strack, D. (2001). Patterns of phenylpropanoids in non-inoculated and potato virus Y-inoculated leaves of transgenic tobacco plants expressing yeast-derived invertase. Phytochemistry 56: 535-541.