請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/90581完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張麗冠 | zh_TW |
| dc.contributor.advisor | Li-Kwan Chang | en |
| dc.contributor.author | 陳聿琳 | zh_TW |
| dc.contributor.author | Yu-Lin Chen | en |
| dc.date.accessioned | 2023-10-03T16:43:34Z | - |
| dc.date.available | 2023-11-09 | - |
| dc.date.copyright | 2023-10-03 | - |
| dc.date.issued | 2023 | - |
| dc.date.submitted | 2023-08-04 | - |
| dc.identifier.citation | Abernathy, E. and B. Glaunsinger (2015). "Emerging roles for RNA degradation in viral replication and antiviral defense." Virology 479-480: 600-608.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/90581 | - |
| dc.description.abstract | EB 病毒 (Epstein-Barr virus, EBV) 是第一個在人類被證實具有致癌性的病毒，感染全球超過九成的人口。EB 病毒的生活史可分成潛伏期 (latency) 與溶裂期 (lytic cycle)，在溶裂期時，病毒的極早期基因 BRLF1 與 BZLF1 所表現的兩個轉錄促進因子 Rta 與 Zta，會進一步活化病毒複製相關的基因表現，促進病毒顆粒產生。此外先前研究已經證實 Rta 能夠促進雙基因 BRLF1-BZLF1 mRNA 中的 Zta 被有效的轉譯。本實驗室先前以免疫沉澱及質譜儀分析發現了一個在病毒進入溶裂期時會與 Rta 結合的人類細胞蛋白質，稱為 meiosis regulator for oocyte development (MIOS)，是與哺乳動物雷帕黴素靶蛋白 （mammalian target of rapamycin, mTOR） 活化相關的上游蛋白質。MIOS 為 GATOR2 (GAP activity towards Rags 2) 複合物中的一員，能夠透過 Rags 蛋白質活化下游的 mTORC1，且在細胞週期與胚胎發育皆扮演了重要的角色。本研究以免疫共沉澱分析以及 GST pull-down assay 證實 Rta 會以其錄活化區 (transactivation domain) 與 MIOS 結合。本研究也發現 MIOS 會透過活化轉譯促進 Rta 蛋白質的表現。亦利用先前所建構的雙基因表現質體 pCMV-RZLuc 進行 luciferase reporter assay，證實了 MIOS 會進一步促進 Rta 活化下游 ORF 的轉譯。此外也進一步以帶有 EB 病毒潛伏的 HEK293(2089) 與P3HR1 細胞證實了 MIOS 會加速 EB 病毒進入溶裂期。綜合以上結果，本研究證實了 MIOS 會與 Rta 結合並促進其蛋白質的表現， MIOS 也能夠促進 EB 病毒進入溶裂期，顯示 MIOS 在 EB 病毒的溶裂期扮演了非常重要的角色。 | zh_TW |
| dc.description.abstract | The Epstein-Barr virus (EBV) is the first virus proven to be oncogenic in humans, infecting over 90% of the global population. The life cycle of EBV can be divided into two stages, latency and the lytic cycle. During the lytic cycle, the virus expresses two transcriptional activators, Rta and Zta, which are encoded by the immediate-early genes BRLF1 and BZLF1, respectively. These two proteins further activate early and late genes, leading to the production of viral particles. In our laboratory, through immunoprecipitation and mass spectrometry analysis, a cellular protein, named as meiosis regulator for oocyte development (MIOS) was found to interact with Rta. MIOS is an upstream protein associated with the activation of the mammalian target of rapamycin complex 1 (mTORC1), which is a central regulator of mammalian metabolism and physiology. MIOS is a member of the GATOR2 complex and plays important roles in the cell cycle and embryonic development. This study used immunoprecipitation and GST pull-down assays to confirm the interaction between Rta and MIOS. Furthermore, MIOS binds to the transactivation domain of Rta. Additionally, MIOS enhances the expression of the Rta protein through translational activation. By using luciferase reporter assay, this study found that MIOS promotes the translation of downstream open reading frames (ORFs) activated by Rta. Remarkly, this study indicated that MIOS accelerates the transition of EBV into the lytic cycle through experiments conducted in HEK293(2089) and P3HR1 cells carrying latent EBV. Taken together, these results demonstrate that MIOS interacts with Rta and enhances its protein expression. Furthermore, MIOS promotes the lytic transition of EBV, indicating its crucial role in EBV life cycle. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2023-10-03T16:43:34Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2023-10-03T16:43:34Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 口試委員審定書 i
致謝 ii 中文摘要 iii Abstract iv 目錄 v 表目錄 vii 圖目錄 viii 第一章 前言 1 1.1 EB 病毒 (Epstein-Barr virus) 1 1.1.1 簡介 1 1.1.2 EB 病毒的基因型與命名 1 1.1.3 病毒的生活史 (life cycle) 2 1.1.4 EB 病毒的溶裂期 (lytic cycle) 4 1.2 EB 病毒的極早期基因 (immediate-early stage) 5 1.2.1 BRLF1-BZLF1 雙基因 mRNA 5 1.2.2 EB 病毒的 Rta 蛋白質 6 1.2.3 Rta 在 BRLF1-BZLF1 雙基因 mRNA 中所扮演的角色 7 1.2.4 Rta 與宿主蛋白質的結合 8 1.3 真核細胞的轉譯 (translation) 9 1.3.1 轉譯 9 1.3.2 真核細胞中轉譯的調控 10 1.3.3 病毒與轉譯的調控 11 1.3.4 Rta 在轉譯中扮演的角色 13 1.4 哺乳動物雷帕黴素靶蛋白 (mammalian target of rapamycin, mTOR) 13 1.4.1 簡介 13 1.4.2 GAP activity towards Rags (GATOR)在mTORC1所扮演的角色 15 1.4.3 mTORC1 與病毒的關聯性 16 1.4.4 人類 MIOS 蛋白質 (human MIOS) 16 1.5 研究動機 17 第二章 材料與方法 18 2.1 細胞株與細胞培養 18 2.2 EB 病毒溶裂期誘導 18 2.3 菌株與細菌培養 18 2.4 質體建構 19 2.5 細菌轉型 (transformation) 20 2.6 質體 DNA 的萃取 20 2.7 轉染作用 (transfection) 20 2.8 電穿孔法 (Electroporation) 21 2.9 細胞裂解液 (lysate) 製備 21 2.10 共免疫沉澱分析 (Co-immunoprecipitation assay, Co-IP) 21 2.11 穀胱甘肽 S-轉移酶沉降分析 22 2.12 西方墨點法 (Western blot) 23 2.13 核質分離 (nuclear/cytosol fractionation) 23 2.14 螢光素酶活性分析 (luciferase reporter assay) 24 2.15 RNA 分析與定量即時反轉錄聚合酶連鎖反應 (qRT-PCR) 24 第三章 結果 26 3.1 MIOS 與 Rta 的結合 26 3.2 MIOS 會與 Rta 的 C 端結合 27 3.3 MIOS 能增加 Rta 的蛋白質表現 27 3.4 MIOS 可能透過促進轉譯增加 Rta 的表現量 28 3.5 MIOS 會促進 EB 病毒進入溶裂期 29 第四章 討論 31 4.1 Rta 透過轉錄活化區域與 MIOS 結合 31 4.2 MIOS 增加 Rta 蛋白質的表現量 32 4.3 MIOS 增強雙基因 mRNA 下游的 ORF 轉譯的能力 33 4.4 MIOS 對於 Rta 的轉譯調節 34 4.5 mTORC1 對於 MIOS 促進 Rta 蛋白質表現量的影響 35 4.6 MIOS 促進 EB 病毒進入溶裂期 36 4.7 結論 37 第五章 圖表 38 參考文獻 53 附錄 77 | - |
| dc.language.iso | zh_TW | - |
| dc.title | 人類 MIOS 蛋白質在 EB 病毒溶裂期的促進 | zh_TW |
| dc.title | Enhancement of Epstein-Barr virus lytic cycle by human MIOS | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 111-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 劉世東;陳美如;吳育騏;李重霈 | zh_TW |
| dc.contributor.oralexamcommittee | Shih-Tung Liu;Mei-Ru Chen;Yu-Chi Wu;Chung-Pei Lee | en |
| dc.subject.keyword | EB 病毒,Rta,human MIOS,轉譯, | zh_TW |
| dc.subject.keyword | Epstein-Barr virus,,Rta,human MIOS,Translation, | en |
| dc.relation.page | 80 | - |
| dc.identifier.doi | 10.6342/NTU202302697 | - |
| dc.rights.note | 同意授權(限校園內公開) | - |
| dc.date.accepted | 2023-08-08 | - |
| dc.contributor.author-college | 生命科學院 | - |
| dc.contributor.author-dept | 生化科技學系 | - |
| dc.date.embargo-lift | 2028-08-03 | - |
| 顯示於系所單位： | 生化科技學系 | |
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