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| ???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
|---|---|---|
| dc.contributor.advisor | 張麗冠 | |
| dc.contributor.author | Chien-Chang Chen | en |
| dc.contributor.author | 陳建彰 | zh_TW |
| dc.date.accessioned | 2021-05-20T20:06:43Z | - |
| dc.date.available | 2011-08-14 | |
| dc.date.available | 2021-05-20T20:06:43Z | - |
| dc.date.copyright | 2009-08-14 | |
| dc.date.issued | 2009 | |
| dc.date.submitted | 2009-08-11 | |
| dc.identifier.citation | Adams A (1987) Replication of latent Epstein-Barr virus genomes in Raji cells. J Virol 61(5): 1743-1746
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/9018 | - |
| dc.description.abstract | EB病毒 (Epstein-Barr virus；EBV)屬於一種人類疱疹病毒，當病毒進入溶裂期會表現出極早期基因BZLF1並轉譯出Zta蛋白質，Zta為一個多功能蛋白質，主要負責調控基因轉錄及將DNA複製相關的蛋白質聚集到複製起始點。為了探討Zta在細胞中是否與其它細胞內蛋白質結合，對EB病毒溶裂期機制造成影響，所以本研究利用His pull-down assay吸附細胞蛋白質並以MALDI-TOF進行蛋白質定序發現人類的ATP-dependent DNA helicase II Ku80會與Zta結合，接著進一步以共同免疫沉澱分析和GST-pull down證實了Zta會與Ku80在細胞內及細胞外結合並一起結合到oriLyt上，並且觀察到Zta以C端區域與Ku80的N端區域結合，也在Ku80缺陷的P3HR1細胞中觀察到EB病毒溶裂期DNA複製的下降，Ku80也會與Zta和EA-D在細胞內結合。此外，利用冷光分析發現Ku80會增加Zta的轉錄活性。綜合以上，Zta與Ku80結合後會進一步調控EB病毒溶裂期複製。 | zh_TW |
| dc.description.abstract | Epstein-Barr virus (EBV) expresses immediate-early transcription factor Zta during the lytic cycle. Zta is a multifunctional protein that regulates gene transcription and recruits a replication complex at the lytic viral origin of replication. To identify cellular proteins that associate with Zta, this study performed His-Zta pull down analysis to precipitate cellular proteins and identified by MALDI-TOF a human ATP-dependent DNA helicase II Ku80 that interacts with Zta. Furthermore, Zta interacts with Ku80 in vivo and in vitro, forming a complex that binds to ZREs at the oriLyt. Furthermore, depleting Ku80 reduces EBV lytic DNA replication, showing the involvement of Ku80 in EBV lytic development. This study shows that the C-terminal domain of Zta interacts with the N- and M-domain of Ku80. Ku80 and Zta also interact with BMRF1 polymerase processivity factor, suggesting that Ku80 facilitates viral lytic DNA replication . In addition, Ku80 enhances the transcriptional activity of Zta. Taken together, this study found Ku80 involves in EBV lytic replication through an interaction with Zta. | en |
| dc.description.provenance | Made available in DSpace on 2021-05-20T20:06:43Z (GMT). No. of bitstreams: 1 ntu-98-R96b47405-1.pdf: 1025601 bytes, checksum: a30a621ba2799f11d24ec230b058fd1c (MD5) Previous issue date: 2009 | en |
| dc.description.tableofcontents | 口試委員會審定書…………………………………………………………………… i
誌謝…………………………………………………………………………………....ii 中文摘要…………………………………………………………………………...... iii 英文摘要…………………………………………………………………………….. iv 前言……………………………………………………………………………………1 一、 EB病毒……………………………………………………………………..1 二、 EB病毒的生活史…………………………………………………………..1 三、 極早期基因BRLF1………………………………………………………...2 四、 極早期基因BZLF1…………………………………………………………3 五、 EB病毒的潛伏期DNA複製………………………………………………4 六、 EB病毒的溶裂期DNA複製………………………………………………5 七、 Ku蛋白質…………………………………………………………………...7 八、 Ku蛋白參與DNA雙股斷裂修補NHEJ路徑……………………………7 九、 Ku蛋白質與DNA轉錄的調控……………………………………………8 十、 Ku蛋白質與DNA複製的活化……………………………………………9 研究目的……………………………………………………………………………..11 實驗方法……………………………………………………………………………..12 結果…………………………………………………………………………………..20 一、 細胞內與Zta結合的蛋白質分析…………………………………………20 二、 Zta與Ku80在細胞內的結合……………………………………………..20 三、 Zta直接與Ku80結合……………………………………………………..21 四、 Zta與Ku80的結合區域分析……………………………………………..21 五、 Ku80與Zta會結合到EB病毒的溶裂期複製起始點的ZRE序列….......22 六、 Ku80與Zta會結合到EB病毒的溶裂期複製起始點 (oriLyt) ………….24 七、 Ku80與BMRF1 polymerase processivity factor在細胞內的交互作用…25 八、 Ku80缺陷時抑制EB病毒溶裂期DNA複製…………………………….25 九、 Ku80活化EB病毒的DNA複製起始…………………………………….26 十、 Ku80強化Zta轉錄因子的活性…………………………………………..27 討論………………………………………………………………….. ……………...29 參考文獻…………………………………………………………………….……….50 附錄…………………………………………………………………………………..64 圖目錄………………………………………………………………………………..35 圖 1：EB病毒的生活史………………………………………………………...35 圖 2：EB病毒的溶裂期………………………………………………………...36 圖 3：EB病毒的溶裂期DNA複製起始區域的序列…………………………..37 圖 4：細胞內與Zta結合的蛋白質……………………………………………...38 圖 5：Zta與Ku80在細胞內的結合…………………………………………….39 圖 6：Zta與Ku80為直接結合………………………………………………….40 圖 7：Ku80與Zta的結合區域分析…………………………………………….41 圖 8：Ku80與Zta會結合到oriLyt的ZRE序列上……………………………..42 圖 9：Ku80與Zta形成複合體結合到oriLyt上………………………………..43 圖 10：Ku80與Zta結合到oriLyt上……………………………………………44 圖 11：Ku80與Zta和BMRF1在細胞內的結合……………………………….45 圖 12：Ku80缺陷的細胞導致EB病毒溶裂期DNA複製量的下降…………..46 圖 13：Ku80促進Zta活化EB病毒溶裂複製的能力………………………….47 圖 14：Ku80促進Zta的轉錄活性能力………………………………………...48 圖15：Ku80與Zta對pBHLF1和pRp-ZRE的影響…………………………….49 表目錄……………………………………………………………………………......64 附表…………………………………………………………………………….. 64 | |
| dc.language.iso | zh-TW | |
| dc.title | Ku80與EB病毒Zta蛋白質的功能研究 | zh_TW |
| dc.title | Ku80 and the Functions of Zta of Epstein-Barr Virus | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 97-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 劉世東,陳美如,張堯,張世宗 | |
| dc.subject.keyword | 人類疱,疹病毒第四型,Zta,Ku80,溶裂期,DNA複製, | zh_TW |
| dc.subject.keyword | Epstein-barr virus,Zta,Ku80,Lytic cycle,DNA replication, | en |
| dc.relation.page | 66 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2009-08-11 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 微生物與生化學研究所 | zh_TW |
| Appears in Collections: | 微生物學科所 | |
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|---|---|---|---|
| ntu-98-1.pdf | 1 MB | Adobe PDF | View/Open |
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