项目名称： La(III)通过衔接蛋白基因异常甲基化激活植物胞吞作用探究

项目编号： No.21501068

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 环境科学、安全科学

项目作者： 王丽红

作者单位： 江南大学

项目金额： 20万元

中文摘要： 稀土在可食用植物中累积并通过食物链蓄积于人体，影响人类健康，已成不争事实。亟需建立可食用植物中稀土限量标准，其首要问题是明晰稀土作用于植物机理。报道显示，La(III)可打破植物叶片长期进化形成的胞吞惰性，激活胞吞作用，然而激活机制尚不明确。衔接蛋白可绑定网格蛋白而启动胞吞作用，通过DNA启动区甲基化影响其基因表达及含量。鉴此，本项目以模式植物拟南芥为代表，采用亚硫氢酸盐测序和独创毛细管电泳-质谱，探讨La(III)诱导衔接蛋白DNA启动区甲基化与代谢分子种类间关系；利用实时荧光定量PCR、Western杂交、透射电镜，探究衔接蛋白异常甲基化对衔接蛋白基因表达、合成量及招募的影响；使用荧光共振能量转移、酵母双杂交、化学计算、激光共聚焦显微镜技术，研究La(III)对衔接蛋白与网格蛋白相互作用方式及其激活胞吞作用的影响。以期阐明La(III)通过衔接蛋白基因异常甲基化激活植物胞吞作用的机制。

中文关键词： 稀土镧离子；植物胞吞作用；衔接蛋白；DNA甲基化；生物效应

英文摘要： It is incontrovertible that the accumulation of rare earth elements (REEs) in edible plants increases the REEs content in human body through food chain, and finally influences human health. Therefore, the limit standard of REEs in edible plants is of urgent need, however, the primary premise, the mechanism of REEs acting plants, is still not clear. It was reported that La(III) can break the sluggishness of endocytosis in plant leaf cells, which is the result of long-time evolution, and activate the endocytosis of plant leaf cells. However, the mechanism of the activation is unknown. Adapter protein can start endocytosis by binding clathrin, and influence the gene express and the content of adaptin through the methylation of DNA promoter. In this project, Arabidopsis thaliana is chosen as the research object. By using bisulfite sequencing PCR and a newly developed capillary electrophoresis-mass spectrum system, we will study the relationship between the La(III)-induced DNA methylation of adapter protein promoter and the variety of metabolism molecules. Moreover, by using real-time fluorescence quantification, Western blotting and transmission electron microscope, we will explore the effects of the aberrant methylation of adapter protein on the gene expression, synthetic amount and recruitment of itself. Furthermore, using fluorescence resonance energy transfer, yeast two-hybrid, computational chemistry and laser scanning confocal microscope, we will investigate the effects of La(III) on the mode of interaction between adapter protein and clathrin, as well as its effect on the activation of endocytosis. This project is expected for illuminating the mechanism of La(III) activating the endocytosis in plant leaf cells via aberrant DNA methylation of adapter protein.

英文关键词： rare earth lanthanum ion;plant endocytosis;adapter protein;DNA methylation;biological effect