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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 李芳仁(Fang-Jen Lee) | |
| dc.contributor.author | Chia-Lun Liu | en |
| dc.contributor.author | 劉嘉倫 | zh_TW |
| dc.date.accessioned | 2021-06-16T05:26:41Z | - |
| dc.date.available | 2019-10-09 | |
| dc.date.copyright | 2014-10-09 | |
| dc.date.issued | 2014 | |
| dc.date.submitted | 2014-08-14 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/56397 | - |
| dc.description.abstract | 腺嘌呤核苷二磷酸核糖化因子(ADP-ribosylation factors, ARFs)在細胞中被認為扮演著調控囊泡運輸的重要角色。在酵母菌的研究中已經知道第三腺嘌呤核苷二磷酸核糖化相似因子(Arl3p)會募集第一腺嘌呤核苷二磷酸核糖化相似因子(Arl1p)，Arl1p可進一步募集下游Imh1p到反式高爾基氏網(trans Golgi network)上並調控高基氏體(Golgi apparatus)的結構與功能。我們實驗室近來的研究發現一種腺嘌呤核苷二磷酸核糖化因子的鳥糞嘌呤核苷酸交換因子(Syt1p)可活化Arl1p並促進Imh1p被募集到高基氏體上，然而目前對於Syt1p的活性調控仍不是很清楚。本研究中我們發現Syt1p的磷酸化對調控Arl1p活化的重要性並進一步探討Syt1p-Arl1p-Imh1p在細胞中參與在囊泡運輸的功能。 | zh_TW |
| dc.description.abstract | ARF-like (ARL) proteins regulate several vesicular trafficking pathways. In yeast, Arl3p recruits Arl1p GTPase, which is specifically required for recruiting golgin protein Imh1p to the trans-Golgi network (TGN), to determine Golgi function and structure. Our previous data reported that Syt1p, a Sec7 domain-containing protein, could promote the activation of Arl1p and recruitment of Imh1p to the TGN. However, the regulation of Syt1p activity toward Arl1p activation remains unclear. In this study, we demonstrate that phosphorylation of Syt1p play an important role in the regulation of Arl1p activation and provide the evidence to support that Syt1p-Arl1p-Imh1p pathway may participate in the vesicular trafficking inside the cells. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T05:26:41Z (GMT). No. of bitstreams: 1 ntu-103-R01448012-1.pdf: 4117358 bytes, checksum: 2cd963c2430fe68b207927f061816f4a (MD5) Previous issue date: 2014 | en |
| dc.description.tableofcontents | 謝誌 I
中文摘要 II Abstract III Contents IV Introduction 1 ADP-ribosylation factor (ARFs) 1 Guanine nucleotide exchange factors (GEF) 3 Arf-like proteins(ARLs) 5 Arl3p-Syt1p-Arl1p-Imh1p signaling pathway. 6 The phosphorylation on Syt1 N-terminal region is important for its GEF activity. 8 Unfolded protein response(UPR) 9 ER stress and trafficking 12 Genetic interactions between Ypt6p and Arl3p-Syt1p-Arl1p-Imh1p pathway. 13 Materials and Methods 15 Yeast two-hybrid analysis 15 Western blotting 15 Yeast transformation 16 HA beads pull-down 18 G-protein pull-down assay 19 UPR stress treatment 20 Results 21 Imh1p localization while tunicamycin treatment. 21 cis, medial and trans – Golgi Morphology while TM treatment. 22 Functions of Imh1p coiled-coil domain are required for the maintenance of Sft2p localization while TM treatment. 24 The increasing punctate Imh1p colocalize with GFP-Sft2p while TM treatment 25 TM treatment could increase the amount of actived Arl1p in cells. 26 The enhancement of phosphorylation on Syt1pS416 while TM treatment is Ire1p dependent. 27 Overexpression of Syt1pS416D is sufficient to result in the increment of punctate Imh1p in cells. 28 Phosphorylation on Serine-416 of Syt1p is required for the punctate pattern of GFP-Sft2p while TM treatment. 29 Overexpression of Syt1pS416D in ire1∆ cells is not sufficient to maintain Sft2p localization while TM treatment. 30 Tunicamycin sensitivity assay 31 Imh1p and Arl1p are able to suppress the mislocalization of GFP-Sft2p in ypt6∆ cells. 32 Syt1pS416D and Arl1p require Imh1p to suppress Sft2p mislocalization in ypt6∆ cells. 34 Syt1pS416D and Arl1p require Imh1p to suppress recycling defect of Snc1p in ypt6∆ cells. 35 Overexpression of Arl1p, Imh1p, Syt1pS416D and conctitutive active Hac1p can suppress temperature growth defect of ypt6Δ. 36 The function of Syt1p-Arl1p-Imh1p pathway in ypt6∆ cells while TM treatment 37 The interaction between Imh1p and Sft2p. 39 Discussion 40 Figures 45 Figure 1. Imh1p localization while TM treatment. 45 Figure 2. cis, medial and trans – Golgi Morphology while TM treatment. 46 Figure 3. Coiled-coil domain of Imp1p is required for the maintenance of GFP-Sft2p localization while TM treatment. 48 Figure 4. The increasing punctate Imh1p colocalize with GFP-Sft2p while TM treatment 49 Figure 5. TM treatment is able to increase the amount of actived Arl1p in cells. 50 Figure 6. The enhancement of phosphorylation on Syt1pS416 while TM treatment is Ire1p dependent. 51 Figure 7. Overexpression of Syt1pS416D is sufficient to result in increment of punctate Imh1p in cells. 52 Figure 8. Phosphorylation on Serine-416 of Syt1p is required for the punctate pattern of GFP-Sft2p while TM treatment. 53 Figure 9. Overexpression of Syt1pS416D in ire1∆ cells is not sufficient to maintain Sft2p localization while TM treatment. 54 Figure 10. Tunicamycin sensitivity assay 55 Figure 11. The model of the enhancement of Syt1p-Arl1p-Imh1p pathway while TM treatment. 56 Figure 12. Imh1p and Arl1p are able to suppress the mislocalization of GFP-Sft2p in ypt6∆ cells. 58 Figure 13. Syt1pS416D and Arl1p require Imh1p to suppress GFP-Sft2p mislocalization in ypt6∆ cells. 59 Figure 14. Syt1pS416D and Arl1p require Imh1p to suppress the recycling defect of Snc1p in ypt6∆ cells. 60 Figure 15. Over-expression of Arl1p, Imh1p, Syt1pS416D and conctitutive active Hac1p can suppress temperature growth defect of ypt6Δ. 61 Figure 16. The overexpression of Syt1pS416D, Arl1p and Imh1p is able to suppress the mislocalization of GFP-Sft2p and GFP-Snc1p in ypt6∆ cells. 62 Figure 17. The function of Syt1p-Arl1p-Imh1p pathway in ypt6∆ cells while TM treatment. 63 Figure 18. The interaction between Imh1p and Sft2p. 64 Tables 65 Table 1. Yeast strains used in this study 65 Table 2. Plasmids used in this study 66 Table 3. Antibodies used in this study 68 References 69 | |
| dc.language.iso | en | |
| dc.subject | 高基氏體 | zh_TW |
| dc.subject | 反式高爾基氏網 | zh_TW |
| dc.subject | Syt1p | zh_TW |
| dc.subject | Arl3p | zh_TW |
| dc.subject | Imh1p | zh_TW |
| dc.subject | Arl1p | zh_TW |
| dc.subject | Arl3p | zh_TW |
| dc.subject | Arl1p | zh_TW |
| dc.subject | ARFs | zh_TW |
| dc.subject | Imh1p | zh_TW |
| dc.subject | Syt1p | zh_TW |
| dc.subject | 反式高爾基氏網 | zh_TW |
| dc.subject | 高基氏體 | zh_TW |
| dc.subject | 囊泡運輸 | zh_TW |
| dc.subject | 囊泡運輸 | zh_TW |
| dc.subject | ARFs | zh_TW |
| dc.subject | Golgi | en |
| dc.subject | ARFs | en |
| dc.subject | Arl3p | en |
| dc.subject | Arl1p | en |
| dc.subject | Imh1p | en |
| dc.subject | Syt1p | en |
| dc.subject | trans-Golgi network | en |
| dc.subject | vesicular trafficking | en |
| dc.subject | ARFs | en |
| dc.subject | Arl3p | en |
| dc.subject | Arl1p | en |
| dc.subject | Imh1p | en |
| dc.subject | Syt1p | en |
| dc.subject | trans-Golgi network | en |
| dc.subject | Golgi | en |
| dc.subject | vesicular trafficking | en |
| dc.title | 酵母菌第一腺嘌呤核苷二磷酸核糖化相似因子之鳥糞嘌呤核苷酸交換因子的磷酸化對促進第一腺嘌呤核苷二磷酸核糖化相似因子活化之研究 | zh_TW |
| dc.title | Functional Characterization of Syt1p Phosphorylation
on Arl1p Activation in Saccharomyces cerevisiae | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 102-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 鄧述諄(Shu-Chun Teng),王昭雯(Chao-Wen Wang) | |
| dc.subject.keyword | ARFs,Arl3p,Arl1p,Imh1p,Syt1p,反式高爾基氏網,高基氏體,囊泡運輸, | zh_TW |
| dc.subject.keyword | ARFs,Arl3p,Arl1p,Imh1p,Syt1p,trans-Golgi network,Golgi,vesicular trafficking, | en |
| dc.relation.page | 73 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2014-08-14 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 分子醫學研究所 | zh_TW |
| 顯示於系所單位： | 分子醫學研究所 | |
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