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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 余明俊(Ming-Jiun Yu) | |
dc.contributor.author | Jheng-Syuan Shao | en |
dc.contributor.author | 邵正玄 | zh_TW |
dc.date.accessioned | 2021-06-08T00:46:36Z | - |
dc.date.copyright | 2015-09-25 | |
dc.date.issued | 2015 | |
dc.date.submitted | 2015-07-29 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17937 | - |
dc.description.abstract | C型肝炎病毒(Hepatitis C virus, HCV)非結構性蛋白質5A (Non-structural protein 5A)是一個磷酸化蛋白質並且有著兩種磷酸化態,包含有:低度(Hypo)以及高度(Hyper)磷酸化態,參與在病毒的複製以及組裝。磷脂醯肌醇-4磷酸酶第三型alpha(PI4KIIIalpha)是宿主中位於內質網的脂質磷酸酶,可以生成phosphatidylinositol 4-phosphate (PI4P)。PI4KIIIalpha已知能夠影響HCV複製中相當重要的NS5A高度磷酸化態。然而PI4KIIIalpha是調控NS5A的哪個磷酸化位點至今未知。本實驗室先前的研究中找到位於NS5A高度磷酸化態的三個磷酸化位點,分別為絲氨酸222、235以及238 (serine 222, 235 and 238)。而我的研究目標是想了解PI4KIIIalpha是否會影響本實驗室發現三個磷酸化位點,進而影響HCV的病毒複製。在被HCV感染的細胞中,PI4KIIIalpha的抑制能夠減少HCV的複製以及S235的磷酸化,而S222以及S238的磷酸化則下降的不明顯。在細胞染色的實驗中,我觀察到磷酸化的S235會與PI4P以及PI(3,4)P2位於相同位置,暗示我們或許會有一個能與phosphoinositides (PI)進行結合的磷酸酶來參與S235的磷酸化。而在HCV感染的細胞中,將AKT這個能與PI進行結合的磷酸酶進行抑制,也能降低S235的磷酸化。因為已知AKT以及NS5A都能與PI(3,4)P2進行結合,我的實驗結果暗示PI4KIIIalpha能磷酸化PI,進而提升PI(3,4)P2的表現量,將AKT與NS5A靠攏在一起並且進行S235的磷酸化。 | zh_TW |
dc.description.abstract | Phosphatidylinositol-4 kinase III alpha (PI4KIIIalpha) is a host ER-resident lipid kinase involved phosphatidylinositol 4 phosphate (PI4P) and phosphatidylinositol 3, 4 bisphosphate (PI(3,4)P2) biosynthesis. PI4KIIIalpha was shown to affect NS5A hyper-phosphorylation that is essential for hepatitis C viral RNA replication; however, the exact phosphorylation sites regulated by PI4KIIIalpha remained unclear. My laboratory identified three serine phosphorylation sites (S222, S235 and S238) all corresponding to NS5A hyper-phosphorylation in HCVcc-infected hepatocarcinoma cells (Huh 7.5.1). The aims of my study are to understand whether PI4KIIIalpha affects any of the three sites to affect HCV RNA replication. In HCVcc-infected cells, PI4KIIIalpha knockdown inhibited HCV replication with a concomitant decrease primarily in S235 phosphorylation. HCVcc infection elevated the levels of PI4P and PI(3,4)P2 that co-localized with S235 phosphorylated NS5A, suggesting a role of a host phosphatidylinositol (PI)-binding kinase in S235 phosphorylation. In HCVcc-infected cells, knockdown of the PI-binding kinase AKT decreased the levels of S235 phosphorylation and HCV RNA. Because both AKT and NS5A bind PI(3,4)P2, my data suggest that PI4KIIIalpha phosphorylates PI, leading to increased PI(3,4)P2 levels that bring AKT to close proximity of NS5A for S235 phosphorylation needed for replication. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T00:46:36Z (GMT). No. of bitstreams: 1 ntu-104-R02442033-1.pdf: 6449793 bytes, checksum: 9199e588412d217b55fbf6b4e5f024e3 (MD5) Previous issue date: 2015 | en |
dc.description.tableofcontents | 摘要------1
Abstract------2 Introduction------3 Materials and Methods------8 Results------14 Alanine mutation at serine 235 of NS5A abolished HCV replication------14 Stable PI4KIIIalpha knockdown prohibited HCV replication in Huh7.5.1 cells------14 PI4KIIIalpha knockdown after HCVcc infection reduced S235 phosphorylation and HCV RNA levels in Huh7.5.1 cells------15 S235 phosphorylated NS5A co-localized with PI4P and PI(3,4)P2 but not with PI4KIIIalpha------16 AKT knockdown reduced S235 phosphorylation in HCVcc-infected Huh7.5.1 cells------18 Discussion------19 Figures and Table------23 References------32 | |
dc.language.iso | en | |
dc.title | 磷脂醯肌醇-4磷酸酶第三型alpha調控C型肝炎病毒非結構性蛋白質NS5A絲氨酸235磷酸化與病毒複製 | zh_TW |
dc.title | Phosphatidylinositol-4 Kinase III alpha (PI4KIIIalpha) Regulates HCV NS5A Hyper-Phosphorylation at Serine 235 and Viral Replication | en |
dc.type | Thesis | |
dc.date.schoolyear | 103-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 張明富(Ming-Fu Chang),陳士隆(Steve S.-L. Chen) | |
dc.subject.keyword | C肝病毒,非結構性蛋白質5A,高度磷酸化, | zh_TW |
dc.subject.keyword | hepatitis c virus,NS5A,hyper-phosphorylaion, | en |
dc.relation.page | 34 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2015-07-29 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 生物化學暨分子生物學研究所 | zh_TW |
顯示於系所單位: | 生物化學暨分子生物學科研究所 |
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