酿酒酵母蛋白激酶A催化亚基细胞定位对其功能特异性的调控机制研究

项目名称： 酿酒酵母蛋白激酶A催化亚基细胞定位对其功能特异性的调控机制研究

项目编号： No.31301536

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 董健

作者单位： 天津科技大学

项目金额： 23万元

中文摘要： 研究酿酒酵母蛋白激酶A催化亚基细胞定位对其功能特异性的调控机制,既有助于完善蛋白激酶A对调控细胞周期和胁迫响应机的生理功能，又可为高耐受性酵母工业菌种分子育种提供理论指导，具有重要的理论意义和应用价值。本项目拟从三个方面探索催化亚基蛋白定位对细胞生长和胁迫耐受性调控机制。首先从分析各催化亚基分别对细胞生长与胁迫耐受性的特异性调控作用出发，发现各亚基调控细胞生长和耐受性的分工情况；其次，通过绿色荧光蛋白标签及其成像技术，观察游离态以及环腺苷酸信号通路激活过程中各催化亚基胞内的静态定位以及动态追踪，研究各催化亚基自身胞内定位属性，以及在激活过程中胞内动态迁移的差异性；最后，以fil1突变株为材料，揭示细胞生长和耐受性矛盾双方相互协调的分子机制，建立蛋白激酶A催化亚基胞内定位对其功能之间的关联, 从分子水平上阐述蛋白激酶A催化亚基分别对细胞生长速率和胁迫耐受性的特异性调控作用和调控机制。

中文关键词： 催化亚基；腺苷酸环化酶；耐受性；信号通路；

英文摘要： This study mainly focuses on the mechanism of protein localization of Protein Kinase A catalytic subunits in regulating their distinct functions in the yeast Saccharomyces cerevisiae. The results will not only be helpful to understand the regulation mechanism of Protein Kinase A in functioning on cell growth and stress resistances, but also with theoretical importance for the breeding of industrial strains with improved stress resistances.In this work, we will uncover the regulation mechanism of Tpk localization in its functions via the following three points: Firstly, we will study the effect of Tpks in regulating cell growth rates and stress resistances, revealing the distinct biological functions of Tpks; Secondly, through the GFP tagged with Tpks, we will trace the localization of Tpks in cells released from the negative regulation of Bcy1 regulatory subunit, as well as in glucose-induced cells, whose cAMP/PKA signalling was acivated by glucose signals. This part of work will find the distinctions of Tpk subcellular localization. Finally, using fil1 mutant as the background, we will study the molecular mechanism of Tpks in regualting cell growth and stress resistance, which are incompatible in wildtype cells, exploring the connection between the Tpks subcellular localization and their divers functions, and f

英文关键词： catalytic subunit；adenylate cyclase；stress tolerance；signalling pathway；