ATG基因调控酵母自噬的系统研究

发布时间：2018-03-29 18:07

本文选题：细胞自噬　切入点：酵母　出处：《华中科技大学》2016年硕士论文

【摘要】：细胞自噬(Autophagy)是高度保守的生物过程,是细胞维持完整性、内稳态和存活所必需的。细胞自噬主要有三种形式:巨自噬(Macroautophagy)、微自噬(Microautophagy)和分子伴侣介导的自噬(Chaperone-mediated autophagy,CMA),我们在本论文中提到的自噬单指巨自噬。自噬异常与许多种疾病相关。因此,自噬分子机制的深入研究非常必要。哺乳动物中自噬相关基因在酵母中有相应的同源物,说明自噬过程高度保守。因此,酵母中的自噬研究可为哺乳动物中自噬研究提供新的思路。目前,在酵母中已经发现40个自噬相关基因(AuTophaGy-related gene,ATG)。酵母(Yeast)具有繁殖快、培养方便和遗传操作简单等特点,许多实验可以在酵母中开展来研究自噬发生各个阶段。因此,本研究选择酵母作为模式生物进行自噬相关研究。本工作以酿酒酵母为研究对象,在减氮饥饿条件下,通过观察0-4h内GFP-ATG8在酵母液泡内外的动态变化,对28个ATG基因敲除菌进行分析并分类,同时对减氮自噬关键ATG蛋白质磷酸化进行生物信息学分析。减氮处理后,根据实验结果表型将减氮自噬相关基因分为两类:减氮自噬关键基因和减氮自噬无影响基因。减氮自噬相关基因分类后,本工作根据酵母蛋白质磷酸化位点数据,整理出18个ATG蛋白质的125个磷酸化位点,用iGPS软件预测自噬基因表达蛋白质位点特异性激酶,共得出13个ATG蛋白的53个蛋白激酶,并构建ATG蛋白质及其激酶网络。未来,本研究工作将对减氮自噬ATG蛋白质的功能磷酸化位点进行实验验证,同时,对减氮自噬关键ATG蛋白质及其激酶的功能关系进行研究。希望这些研究工作能为今后高等真核生物中自噬分子机制研究提供新的方法和思路。
[Abstract]:Autophagy is a highly conserved biological process that maintains cell integrity. Necessary for homeostasis and survival. Autophagy occurs in three main forms: Macroautophagyus, microautophagyi, and chaperone-mediated autophagy.Anophonic autophagy, referred to in this paper, is associated with many diseases. It is necessary to further study the mechanism of autophagy. The homologues of autophagy related genes in mammalian yeast indicate that autophagy is highly conserved. The study of autophagy in yeast can provide a new idea for the study of autophagy in mammals. At present, 40 autophagy related genes have been found in yeast, such as fast propagation, easy culture and simple genetic operation. Many experiments can be carried out in yeast to study the various stages of autophagy. By observing the dynamic changes of GFP-ATG8 in and out of yeast vacuoles within 0-4 h, 28 ATG knockout bacteria were analyzed and classified, and the phosphorylation of key ATG proteins in nitrogen-reducing autophagy was analyzed by bioinformatics. According to the phenotype of the experiment, the genes associated with nitrogen reduction autophagy were divided into two categories: the key genes of nitrogen reduction autophagy and the genes with no effect on nitrogen reduction autophagy. After the classification of the genes associated with nitrogen reduction autophagy, the data of phosphorylation sites of yeast protein were used in this work. A total of 125 phosphorylation sites of 18 ATG proteins were sorted out, and 53 protein kinases of 13 ATG proteins were obtained using iGPS software to predict protein site-specific kinases expressed in autophagy genes, and the ATG protein and its kinase network were constructed. In this study, the functional phosphorylation sites of nitrogen-reducing autophagy ATG protein were verified. The functional relationships of key ATG proteins and their kinases in nitrogen-reducing autophagy were studied. It is hoped that these studies will provide new methods and ideas for the study of autophagy molecular mechanism in higher eukaryotes in the future.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q933

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