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標題: | 酵母菌第一及第三腺嘌呤核苷二磷酸核醣化因子相似蛋
白功能性探討 Functional Characterization of ARF-like protein Arl1p and Arl3p |
作者: | Yi-Jie Chen 陳怡潔 |
指導教授: | 李芳仁(Fang-Jen Lee) |
關鍵字: | Arl3p,Arl1p,囊泡運輸,反式高爾基體網絡,細胞質至液胞傳遞途徑,負回饋, Arl3p,Arl1p,vesicle transport,TGN,Cvt pathway,feedback regulation, |
出版年 : | 2016 |
學位: | 碩士 |
摘要: | 腺嘌呤核苷二磷酸核醣化因子相似蛋白 (ARL) 是腺嘌呤核苷二磷酸核醣化因子 (ADP-ribosylation factors , ARF) 家族的一員,其功能主要參與在反氏高爾基氏體中囊泡運輸的過程。在酵母菌中,已知活化態之第三腺核苷二磷酸醣化因子相似蛋白(Arl3p)可以招集並活化第一腺核苷二磷酸醣化因子相似蛋白(Arl1p)到反氏高爾基氏體上執行功能。目前研究已經知道第一腺核苷二磷酸醣化因子相似蛋白(Arl1p)參與在三條路徑中,分別是:吸引Imh1p 和Gga2p 到反氏高爾基氏體上,及運輸Gas1p 到細胞膜上。在先前研究顯示Arl3p 為Arl1p 上游蛋白質,然而我們目前對於Arl3p 之生理功能不清楚且其互相調控機制亦尚未明瞭。本研究利用酵母菌作為模式生物來釐清Arl3p 之生理功能及Arl3p 如何去調控下游蛋白質Arl1p。
在我們的實驗中發現,Arl3p-Arl1p 這條路徑會協同Cog8p 參與在細胞質至液胞傳遞途徑(cytoplasm-to-vacuole targeting pathway, Cvt pathway),並且發現Arl3p-Arl1p 是藉由影響到Atg9p 的正向運輸而導致細胞質至液胞傳遞途徑的缺陷。除此之外,我們也證實Arl3p 與Arl1p 之間存在直接的交互作用, 並且初步利用點菌實驗發現可能在Arl3p 與 Arl1p 中藉由交互作用存在一個負回饋的調控機制。 ARF-like proteins (ARL) are members of ADP-ribosylation factors (ARF) family of small guanine-nucleotide-binding proteins. ARLs participate in the vesicle trafficking at the trans-Golgi network (TGN). In yeast, activated Arl3p recruits Arl1p to the TGN. It is known that Arl1p participates in three different pathways at the TGN, including recruitment of golgin Imh1p, daptor protein Gg2p to the Golgi and transport of glycosylphosphatidylinositol (GPI)-anchored protein Gas1p to plasma membrane. In previous studies, we know that Arl3p is the upstream regulator of Arl1p. However, the biological function of Arl3p and how the Arl3p regulates Arl1p is still unclear. In this study, we show that Arl3p-Arl1p pathway plays a role in cytoplasm to vacuole targeting (Cvt) pathway in coordination with cog8p. Arl1p and Arl3p can regulate Atg9p anterograde transport from TGN to pre-autophagosomal structure (PAS) to promote the Cvt vesicle formation. On the other hand, in vivo and in vitro experiments indicate that active form Arl3p interacts with active form Arl1p. Furthermore, we found that the direct interaction between Arl3p and Arl1p may be a potential regulation mechanism for Arl1p downstream function. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/49555 |
DOI: | 10.6342/NTU201602957 |
全文授權: | 有償授權 |
顯示於系所單位: | 分子醫學研究所 |
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