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| ???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
|---|---|---|
| dc.contributor.advisor | 李芳仁 | |
| dc.contributor.author | Chia-Lu Wu | en |
| dc.contributor.author | 巫佳儒 | zh_TW |
| dc.date.accessioned | 2021-07-10T21:52:37Z | - |
| dc.date.available | 2021-07-10T21:52:37Z | - |
| dc.date.copyright | 2019-08-28 | |
| dc.date.issued | 2019 | |
| dc.date.submitted | 2019-08-14 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77247 | - |
| dc.description.abstract | 從酵母菌乃至哺乳類動物,腺嘌呤核苷二磷酸核糖化相似因子 (Arf-like protein,Arl) 在細胞體內扮演著調控囊泡運輸的功能。而在釀酒酵母中,第一腺嘌 呤核苷二磷酸核糖化相似因子 (Arl1) 負責上招募高爾基體蛋白 Imh1 至反式高爾 基氏體網路 (trans-Golgi network)已為人所知;然而,其下游的運輸貨物以及功能 在先前的報導中並未獲得詳盡的研究。在我們先前的研究中發現,當細胞遭遇未折 疊蛋白反應時,細胞會促進 Arl1 以及其下游 Imh1 的活化,但是當時我們並未探 究其中作用的機轉以及 Arl1 和 Imh1 被活化的目的。在本篇研究中,我們發現當 細胞遭遇未折疊蛋白反應時,被活化的 Arl1 以及 Imh1 會 Sft2 一起調控細胞中的 囊泡運輸。另外,我們發現這樣的機制與 GARP 所主導的反向運輸無關。反之, 我們的研究指出,Imh1 和 Sft2 會一起調控 v-SNARE 蛋白 Snc1 的正向運輸。當細 胞裡缺乏 Imh1 或是 Sft2 時,Snc1 會卡在反式高爾基氏體網路而無法送達至細胞 膜。總結本篇,我們的研究意謂著細胞囊泡運輸系統在內質網壓力下的重要性。更 多的是,我們找到了當細胞遭遇未折疊蛋白反應時,Imh1 以及 Sft2 在囊泡運輸的功能。 | zh_TW |
| dc.description.abstract | ADP-ribosylation factors-like protein 1 (Arl1) is an important regulator of intracellular vesicular trafficking in organisms ranging from yeast to mammals. In Saccharomyces cerevisiae, Arl1 recruits its effector Imh1 to the trans-Golgi network (TGN). However, the downstream cargo or function of Arl1-Imh1 is still unclear. According to previous studies, Unfolded-Protein Response (UPR) promotes the activation of Arl1 and thus recruits Imh1 to the Golgi, but its downstream effect upon UPR remains to be elucidated. In this study, we validate that activated Imh1 recruits Sft2 to the TGN, which is involved in vesicular transport when cells confront UPR. Moreover, we found out that this regulation is independent of GARP complex recruitment, a complex important to retrograde trafficking. In contrast, Imh1 and Sft2 mediate the anterograde of v-SNARE protein Snc1 that is stuck in TGN in the absence of Imh1 or Sft2 upon UPR. Together, our study demonstrates the specific function of golgin Imh1 and Sft2 in trafficking machinery when cells encounter UPR. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-10T21:52:37Z (GMT). No. of bitstreams: 1 ntu-108-R06448003-1.pdf: 1796312 bytes, checksum: 9314412ca901275fb723a23da8b0673a (MD5) Previous issue date: 2019 | en |
| dc.description.tableofcontents | 致謝 .............................................................................................................. 2
中文摘要 ...................................................................................................... 6 Abstract ....................................................................................................... 7 Introduction................................................................................................. 8 ADP-ribosylation Factors .......................................................................... 8 Material and Methods .............................................................................. 18 Tables ......................................................................................................... 28 Table 1. Yeast strains used in this study ................................................................. 28 Table 2. Plasmids used in this study ....................................................................... 30 Table 3. Antibodies used in this study .................................................................... 32 Results........................................................................................................ 33 Part I: Functional Characterization of Sft2 upon UPR in Saccharomyces cerevisiae. ................................................................................................................................ 33 The enhanced-recruitment of Imh1 is required for Sft2 localization upon UPR. ........................................................................................................................ 33 Sft2 acts downstream of Imh1 to regulate SNARE proteins trafficking upon UPR. ............................................................................................................... 34 Imh1-Sft2 regulate the anterograde transport of Snc1 upon UPR.................. 35 The anterograde pathway upon UPR is independent of GARP complex. ...... 37 The N-terminus of Sft2 might be important to regulating v-SNARE transport upon UPR. ...................................................................................................... 38 Got1 is also involved in v-SNARE anterograde transport upon UPR............ 39 Got1 might participate in the pathway indirectly upon UPR. ........................ 40 Part II: Identification of an arginine permease Can1 as a putative cargo of Arl1 small GTPase. ......................................................................................................... 41 Arl3-Arl1 cascade may be involved in Can1 transport through Syt1 independent pathway. ..................................................................................... 41 Arl1 might regulate Can1 through its anterograde transport. ......................... 43 Other Arl1-related proteins also mediate Can1 transport. .............................. 44 Drs2 might regulate Can1 together with Arl1. ............................................... 45 Discussion .................................................................................................. 47 Part I: Functional Characterization of Sft2 upon UPR in Saccharomyces cerevisiae ................................................................................................................................ 47 The role of Sft2 in mediating anterograde trafficking upon UPR. ................. 47 The differential role of Imh1 and Sft2 in suppression of ypt6∆ cells and upon UPR. ............................................................................................................... 48 The involvement of Got1 in vesicular trafficking mediated by Imh1-Sft2 upon UPR. ............................................................................................................... 49 Part II: Identification of an arginine permease Can1 as a putative cargo of Arl1 small GTPase .......................................................................................................... 50 Cargos that is also regulated by Arl1-Arl3. .................................................... 51 Figures ....................................................................................................... 52 Figure 1. UPR facilitates the elevated-recruitment of Imh1 which is colocalized with GFP-Sft2.............................................................................. 52 Figure 2. Imh1 is required for the maintenance of GFP-Sft2 localization upon UPR. ............................................................................................................... 53 Figure 3. Sft2 acts downstream of Imh1 upon UPR....................................... 54 Figure 4. Sft2 is required for proper trafficking of Snc1 and Tlg1 upon UPR. ........................................................................................................................ 55 Figure 5. Imh1 and Sft2 regulate Snc1 in anterograde trafficking upon UPR. ........................................................................................................................ 57 Figure 6. Snc1 shows different mislocalized pattern in various recycling mutant strains.................................................................................................. 59 Figure 7. Localization of GARP complex is not affected by Sft2 or Imh1 upon UPR. ............................................................................................................... 60 Figure 8. Imh1 interact with Sft2 in vitro. ...................................................... 61 Figure 9. Imh1 interact with Sft2 in vitro. ...................................................... 63 Figure 10. N-terminus of Sft2 might be indispensable for its function upon UPR. ............................................................................................................... 66 Figure 11. Got1 is also required for Snc1 but not Tlg1 trafficking upon UPR. ........................................................................................................................ 67 | |
| dc.language.iso | en | |
| dc.subject | 高爾基氏體 | zh_TW |
| dc.subject | 第一腺嘌呤核?二磷酸核糖化相似因子 | zh_TW |
| dc.subject | 高爾機體蛋白 | zh_TW |
| dc.subject | 未折疊蛋白反應 | zh_TW |
| dc.subject | 正向運輸 | zh_TW |
| dc.subject | UPR | en |
| dc.subject | Arl1 | en |
| dc.subject | Imh1 | en |
| dc.subject | Sft2 | en |
| dc.subject | anterograde trafficking | en |
| dc.title | 第一腺嘌呤核苷二磷酸核醣化因子相似蛋白於酵母菌
之功能性探討 | zh_TW |
| dc.title | Functional Characterization of ADP-Ribosylation Factor- Like Protein 1 in Saccharomyces cerevisiae | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 107-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 鄧述諄,陳瑞華,林敬哲,王昭雯 | |
| dc.subject.keyword | 高爾基氏體,第一腺嘌呤核?二磷酸核糖化相似因子,高爾機體蛋白,未折疊蛋白反應,正向運輸, | zh_TW |
| dc.subject.keyword | Arl1,Imh1,Sft2,UPR,anterograde trafficking, | en |
| dc.relation.page | 91 | |
| dc.identifier.doi | 10.6342/NTU201903594 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2019-08-14 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 分子醫學研究所 | zh_TW |
| Appears in Collections: | 分子醫學研究所 | |
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| ntu-108-R06448003-1.pdf Restricted Access | 1.75 MB | Adobe PDF |
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