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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 林詩舜(Shih-Shun Lin) | |
| dc.contributor.author | Phuong Anh Tran | en |
| dc.contributor.author | 陳芳英 | zh_TW |
| dc.date.accessioned | 2022-11-24T03:34:42Z | - |
| dc.date.available | 2026-08-03 | |
| dc.date.available | 2022-11-24T03:34:42Z | - |
| dc.date.copyright | 2021-08-11 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-08-05 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81180 | - |
| dc.description.abstract | Potyvirus 的基因靜默抑制子 HC-Pro 在抑制轉錄後基因靜默作用 (PTGS) 中起著重要的作用。然而，目前對於HC-Pro如何抑制微型核酸 (miRNA) 甲基化與 ARGONAUTE 1 (AGO1) 的加载以形成 RISC複合體 (RISC)的作用途徑尚未清楚。我们先前的研究表明HC-Pro 利用自噬途徑中的相關蛋白ATG8a介導降解ARGONAUTE1 (AGO1) 蛋白並抑制HEN1甲基轉移酶活性。本研究利用蛋白的螢光共位 (colocalization) 和福斯特能量共振轉移 (FRET) 研究in vivo細胞內蛋白質與蛋白質相互作用。我們的結果顯示 HC-Pro 与 AGO1 在Suppression body (S-body) 有間接相互作用。另外HC-Pro 和 ATG8a 有colocalization現象和相互作用，再次證明ATG8a 與HC-Pro 間接誘導 AGO1 降解有關。FRET 結果也顯示HC-Pro直接物理相互作用抑制 HEN1 甲基轉移酶活性，而未甲基化的微型核酸 (unMet-miRNA) 無法加載至 RISC 复合體。除此之外，HC-Pro 與Processing body (P-body) 標記物的mRNA-去頭蓋蛋白質2 (DCP2)有colocalization現象和相互作用。實驗結果顯示HC-Pro 在細胞質形成S-body。根據PTGS 分子如AGO1、HEN1、ATG8a 和 DCP2 的焦距大小結果，我們推測S-body會吸引這些蛋白進入。了解 S-body 和 PTGS 分子之間的關係讓我们更加理解 HC-Pro 如何抑制微型核酸途徑，并確認微型核酸被甲基化與加載到 AGO1 的位置。這些研究可闡明HEN1 如何喪失甲基化活性。進一步的研究可以顯示未甲基化的微型核酸如何誘導ATG8a自噬蛋白介導 AGO1 降解。綜合來說，這項研究有助于理解 PTGS 調控與植物和病毒之間的相互作用。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-24T03:34:42Z (GMT). No. of bitstreams: 1 U0001-0308202117144900.pdf: 2185090 bytes, checksum: 61d46758d289b2470ebec4dd59d1ff99 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | "Acknowledgement II 中文摘要 III Abstract IV Contents VI List of figures VIII Introduction 1 PTGS pathway and the viral suppressor HC-Pro 1 TuGR and TuGK 3 FRET application in protein-protein interaction research 3 Materials and Methods 5 Plant materials and growth conditions 5 pENTR clone construction 5 Binary vector construction 6 Transient expression by agro-infiltration 7 Confocol microscopy 7 Size measurement 8 Super-resolution images analysis 8 Fluorescence Resonance Energy Transfer (FRET) 8 Results 10 Subcellular localization of HC-Pro suppressor, and components of gene silencing and autophagy 10 Evaluation of the size of different bodies 10 Subcellular co-localization of HC-Pro and HEN1 11 Subcellular co-localization of HC-Pro and DCP2 11 Subcellular co-localization of HC-Pro and AGO1 12 Subcellular co-localization of HC-Pro and ATG8a 13 Evaluation of the physical interaction between HC-Pro and gene silencing components by FRET 14 Super-resolution images of S-body 14 Discussion 16 The S-body is an important place for HC-Pro to inhibit miRNA pathway 16 Physical interaction between HC-Pro and certain RNA silencing components 17 HC-ProK versus HC-ProR 19 Using ultra-resolution microscope to study the composition of S-body 19 The working hypotheses 20 The future perspective 22 Conclusion 24 References 25 Figures 31" | |
| dc.language.iso | en | |
| dc.subject | 轉錄後基因靜默作用 | zh_TW |
| dc.subject | 基因靜默抑制子 | zh_TW |
| dc.subject | 蛋白质-蛋白质相互作用 | zh_TW |
| dc.subject | DCP2 | en |
| dc.subject | Protein-protein interaction | en |
| dc.subject | TuMV | en |
| dc.subject | HC-Pro | en |
| dc.subject | PTGS | en |
| dc.subject | AGO1 | en |
| dc.subject | ATG8a | en |
| dc.subject | HEN1 | en |
| dc.subject | S-body | en |
| dc.title | 利用細胞生物學策略分析 HC-Pro 如何作用並干擾重要核酸靜默反應元件之研究 | zh_TW |
| dc.title | Investigation of the HC-Pro interaction and interfering with critical components of RNA silencing through cell biology approach | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 吳素幸(Hsin-Tsai Liu),邱子珍(Chih-Yang Tseng),陳荷明,朱士維 | |
| dc.subject.keyword | 蛋白质-蛋白质相互作用,基因靜默抑制子,轉錄後基因靜默作用, | zh_TW |
| dc.subject.keyword | Protein-protein interaction,TuMV,HC-Pro,PTGS,AGO1,ATG8a,DCP2,HEN1,S-body, | en |
| dc.relation.page | 48 | |
| dc.identifier.doi | 10.6342/NTU202102054 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-08-06 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 生物科技研究所 | zh_TW |
| dc.date.embargo-lift | 2026-08-03 | - |
| 顯示於系所單位： | 生物科技研究所 | |
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