本文選題：細(xì)胞自噬　切入點：酵母　出處：《華中科技大學(xué)》2016年碩士論文

【摘要】：細(xì)胞自噬(Autophagy)是高度保守的生物過程,是細(xì)胞維持完整性、內(nèi)穩(wěn)態(tài)和存活所必需的。細(xì)胞自噬主要有三種形式:巨自噬(Macroautophagy)、微自噬(Microautophagy)和分子伴侶介導(dǎo)的自噬(Chaperone-mediated autophagy,CMA),我們在本論文中提到的自噬單指巨自噬。自噬異常與許多種疾病相關(guān)。因此,自噬分子機(jī)制的深入研究非常必要。哺乳動物中自噬相關(guān)基因在酵母中有相應(yīng)的同源物,說明自噬過程高度保守。因此,酵母中的自噬研究可為哺乳動物中自噬研究提供新的思路。目前,在酵母中已經(jīng)發(fā)現(xiàn)40個自噬相關(guān)基因(AuTophaGy-related gene,ATG)。酵母(Yeast)具有繁殖快、培養(yǎng)方便和遺傳操作簡單等特點,許多實驗可以在酵母中開展來研究自噬發(fā)生各個階段。因此,本研究選擇酵母作為模式生物進(jìn)行自噬相關(guān)研究。本工作以釀酒酵母為研究對象,在減氮饑餓條件下,通過觀察0-4h內(nèi)GFP-ATG8在酵母液泡內(nèi)外的動態(tài)變化,對28個ATG基因敲除菌進(jìn)行分析并分類,同時對減氮自噬關(guān)鍵ATG蛋白質(zhì)磷酸化進(jìn)行生物信息學(xué)分析。減氮處理后,根據(jù)實驗結(jié)果表型將減氮自噬相關(guān)基因分為兩類:減氮自噬關(guān)鍵基因和減氮自噬無影響基因。減氮自噬相關(guān)基因分類后,本工作根據(jù)酵母蛋白質(zhì)磷酸化位點數(shù)據(jù),整理出18個ATG蛋白質(zhì)的125個磷酸化位點,用iGPS軟件預(yù)測自噬基因表達(dá)蛋白質(zhì)位點特異性激酶,共得出13個ATG蛋白的53個蛋白激酶,并構(gòu)建ATG蛋白質(zhì)及其激酶網(wǎng)絡(luò)。未來,本研究工作將對減氮自噬ATG蛋白質(zhì)的功能磷酸化位點進(jìn)行實驗驗證,同時,對減氮自噬關(guān)鍵ATG蛋白質(zhì)及其激酶的功能關(guān)系進(jìn)行研究。希望這些研究工作能為今后高等真核生物中自噬分子機(jī)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.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：Q933

【相似文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 衛(wèi)亞平;ATG基因調(diào)控酵母自噬的系統(tǒng)研究[D];華中科技大學(xué);2016年

，

本文編號：1682248