請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23382
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 施嘉和(Chiaho Shih) | |
dc.contributor.author | Er-Yi Huang | en |
dc.contributor.author | 黃爾毅 | zh_TW |
dc.date.accessioned | 2021-06-08T05:00:12Z | - |
dc.date.copyright | 2010-09-09 | |
dc.date.issued | 2010 | |
dc.date.submitted | 2010-08-17 | |
dc.identifier.citation | References 1. Blumberg BS (1997) Hepatitis B virus, the vaccine, and the control of primary cancer of the liver. Proc Natl Acad Sci U S A 94: 7121-7125. 2. Purcell RH (1994) Hepatitis viruses: changing patterns of human disease. Proc Natl Acad Sci U S A 91: 2401-2406. 3. Ganem D, Prince AM (2004) Hepatitis B virus infection--natural history and clinical consequences. N Engl J Med 350: 1118-1129. 4. Chang MH, Chen CJ, Lai MS, Hsu HM, Wu TC, et al. (1997) Universal hepatitis B vaccination in Taiwan and the incidence of hepatocellular carcinoma in children. Taiwan Childhood Hepatoma Study Group. N Engl J Med 336: 1855-1859. 5. Liaw YF, Chu CM (2009) Hepatitis B virus infection. Lancet 373: 582-592. 6. Zlotnick A, Cheng N, Conway JF, Booy FP, Steven AC, et al. (1996) Dimorphism of hepatitis B virus capsids is strongly influenced by the C-terminus of the capsid protein. Biochemistry 35: 7412-7421. 7. Zlotnick A, Cheng N, Stahl SJ, Conway JF, Steven AC, et al. (1997) Localization of the C terminus of the assembly domain of hepatitis B virus capsid protein: implications for morphogenesis and organization of encapsidated RNA. Proc Natl Acad Sci U S A 94: 9556-9561. 8. Newman M, Suk FM, Cajimat M, Chua PK, Shih C (2003) Stability and morphology comparisons of self-assembled virus-like particles from wild-type and mutant human hepatitis B virus capsid proteins. J Virol 77: 12950-12960. 9. Le Pogam S, Chua PK, Newman M, Shih C (2005) Exposure of RNA templates and encapsidation of spliced viral RNA are influenced by the arginine-rich domain of human hepatitis B virus core antigen (HBcAg 165-173). J Virol 79: 1871-1887. 10. Newman M, Chua PK, Tang FM, Su PY, Shih C (2009) Testing an electrostatic interaction hypothesis of hepatitis B virus capsid stability by using an in vitro capsid disassembly/reassembly system. J Virol 83: 10616-10626. 11. Chua PK, Tang FM, Huang JY, Suen CS, Shih C (2010) Testing the balanced electrostatic interaction hypothesis of hepatitis B virus DNA synthesis by using an in vivo charge rebalance approach. J Virol 84: 2340-2351. 12. Chu CM, Liaw YF (1987) Intrahepatic distribution of hepatitis B surface and core antigens in chronic hepatitis B virus infection. Hepatocyte with cytoplasmic/membranous hepatitis B core antigen as a possible target for immune hepatocytolysis. Gastroenterology 92: 220-225. 13. Yeh CT, Liaw YF, Ou JH (1990) The arginine-rich domain of hepatitis B virus precore and core proteins contains a signal for nuclear transport. J Virol 64: 6141-6147. 14. Kann M, Sodeik B, Vlachou A, Gerlich WH, Helenius A (1999) Phosphorylation-dependent binding of hepatitis B virus core particles to the nuclear pore complex. J Cell Biol 145: 45-55. 15. Eckhardt SG, Milich DR, McLachlan A (1991) Hepatitis B virus core antigen has two nuclear localization sequences in the arginine-rich carboxyl terminus. J Virol 65: 575-582. 16. Liao W, Ou JH (1995) Phosphorylation and nuclear localization of the hepatitis B virus core protein: significance of serine in the three repeated SPRRR motifs. J Virol 69: 1025-1029. 17. Porterfield JZ, Dhason MS, Loeb DD, Nassal M, Stray SJ, et al. (2010) Full-length hepatitis B virus core protein packages viral and heterologous RNA with similarly high levels of cooperativity. J Virol 84: 7174-7184. 18. Nassal M (1992) The arginine-rich domain of the hepatitis B virus core protein is required for pregenome encapsidation and productive viral positive-strand DNA synthesis but not for virus assembly. J Virol 66: 4107-4116. 19. Hung-Cheng Li, Er-Yi Huang, Pei-Yi Su, Szu-Yao Wu, Ching-Chun Yang, Young-Sun Lin, Wen-Chang Chang, and Chiaho Shih (2010) Nuclear Export and Import of Human Hepatitis B Virus Capsid Protein and Particles. manuscript under revision. 20. Chang J, Sigal LJ, Lerro A, Taylor J (2001) Replication of the human hepatitis delta virus genome Is initiated in mouse hepatocytes following intravenous injection of naked DNA or RNA sequences. J Virol 75: 3469-3473. 21. Yang PL, Althage A, Chung J, Chisari FV (2002) Hydrodynamic injection of viral DNA: a mouse model of acute hepatitis B virus infection. Proc Natl Acad Sci U S A 99: 13825-13830. 22. Yeh CT, Wong SW, Fung YK, Ou JH (1993) Cell cycle regulation of nuclear localization of hepatitis B virus core protein. Proc Natl Acad Sci U S A 90: 6459-6463. 23. Harreman MT, Kline TM, Milford HG, Harben MB, Hodel AE, et al. (2004) Regulation of nuclear import by phosphorylation adjacent to nuclear localization signals. J Biol Chem 279: 20613-20621. 24. Zhang F, White RL, Neufeld KL (2000) Phosphorylation near nuclear localization signal regulates nuclear import of adenomatous polyposis coli protein. Proc Natl Acad Sci U S A 97: 12577-12582. 25. Tagawa T, Kuroki T, Vogt PK, Chida K (1995) The cell cycle-dependent nuclear import of v-Jun is regulated by phosphorylation of a serine adjacent to the nuclear localization signal. J Cell Biol 130: 255-263. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23382 | - |
dc.description.abstract | B型肝炎病毒(HBV)的核心蛋白質(HBc) 在蛋白質序列147-183的carboxyl端上包含一塊富有arginine的區域(ARD)。在之前的研究,我們證明了ARD與HBc的交通信號有關,包含細胞核定位訊號(NLS)以及細胞核輸出訊號(NES)。在研究不同長度的HBc deletion突變型的實驗中,我們指出在交通信號之間的邊界區域對於影響HBc在細胞內的位置非常重要。在這些交通信號之間的邊界區域有共七個serines和一個threonine。以前的報告已經顯示模仿去磷酸化(dephosphorylation)的突變型S162A和S170A能強烈將HBc導向細胞核,而突變型S155A只是溫和的影響HBc在細胞核堆積。然而突變型S162A和S170A造成HBc在細胞核堆積的現象到底是透過活化NLS或者抑制NES尚不清楚。另外在交通信號之間的邊境區域中,其他的serines或者非serines序列是否調控HBc的交通運輸亦不清楚。在這裡我們的研究指出突變型S162A和S170A都能活化NLS並且同時抑制NES。相對於wild type的HBc,HBc突變型S168A在活化NLS但沒有明顯影響NES的情況下將HBc從細胞質移動到細胞核的趨勢增加200倍。突變型S178A則在強烈的抑制NES和較小程度影響NLS的情況下將HBc從細胞質移動到細胞核的趨勢增加117倍。除了serines和threonines之外,另外一個具有明顯表型的突變P163Q讓超過百分之70的細胞具有HBc在細胞核堆積的現象。這個結果可以在hydrodynamic delivery的老鼠模型中被確認。不過,HBc在細胞核堆積的現象在雙突變型S162D/P163Q中減弱了10 倍,這個結果顯示S162D對HBc的影響超越P163Q。總而言之,我們證明了HBc在細胞內的分佈可以受到NLS、NES及其邊境區域的調節。然而在整個HBc ARD區域中缺乏負電荷(serine phosphorylation)強烈促進HBV核心蛋白質在細胞核堆積。 | zh_TW |
dc.description.abstract | Hepatitis B virus (HBV) core protein (HBc) contains an arginine-rich domain (ARD) at the carboxyl terminus of HBc 147-183. Previously, we demonstrated that ARD is associated with trafficking signals, including nuclear localization signals (NLS) and nuclear export signals (NES). In our studies of serially truncated HBc mutants, we noted that border regions between the trafficking signals are very important in affecting the subcellular localization. There are a total of 7 serines and one threonine at the borders between these trafficking signals. Previous reports have shown that dephosphorylation-mimicking mutations S162A and S170A can strongly target HBc to nucleus, while S155A exhibited only mild effect on nuclear accumulation of HBc. Mechanistically, it is unclear if nuclear import of HBc of mutants S162A and S170A is mediated through activation of NLS or inactivation of NES. Furthermore, it remains unclear if other serine or non-serine residues at the borders could also be subject to modulation of trafficking activity. In our studies here, both S162A and S170A can activate NLS and inactivate NES simultaneously. Relative to wild type HBV, HBc mutant S168A exhibited 200 fold increased tendency to shift HBc from cytoplasm to nucleus by activating NLS, with no apparent effect on NES. Mutant S178A exhibited 117 fold increased tendency to shift HBc from cytoplasm to nucleus by strongly inactivating NES and to a lesser extent on NLS. In addition to serines and threonines, a striking phenotypic effect was observed in mutant P163Q with more than 70 % of cells containing nuclear HBc. This result was confirmed by a mouse model with hydrodynamic delivery. However, double mutant S162D/P163Q exhibited a 10-fold reduction of HBc accumulation in the nucleus, suggesting that S162D is dominant over P163Q. In summary, we demonstrated that the subcellular localization of HBc can be regulated by both NLS and NES. Lack of negative charge (serine phosphorylation) spanning the entire HBc ARD strongly promoted nuclear accumulation of HBV core protein. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T05:00:12Z (GMT). No. of bitstreams: 1 ntu-99-D90448003-1.pdf: 8385758 bytes, checksum: 9786ced4653f5c11cba669555aa7800a (MD5) Previous issue date: 2010 | en |
dc.description.tableofcontents | 目 錄 口試委員會審定書……………………………………………………. I 謝辭……………………………………………………………………. II 中文摘要………………………………………………………………. III 英文摘要………………………………………………………………. IV 第一章 INTRODUCTION…………………………………….. 1 第二章MATERIALS AND METHODS………………..…… 4 第三章 RESULTS………………..………………..………… 10 第四章DISCUSSION………………………………………... 19 參考文獻…………………………………………………….…… 51 | |
dc.language.iso | en | |
dc.title | 分析B型肝炎病毒顆粒在細胞內交通運輸的調控機制 | zh_TW |
dc.title | Dissection of the Modulation Mechanism of Intracellular Trafficking Activity of Human Hepatitis B Virus Capsids | en |
dc.type | Thesis | |
dc.date.schoolyear | 98-2 | |
dc.description.degree | 博士 | |
dc.contributor.coadvisor | 林陽生(Young-Sun Lin) | |
dc.contributor.oralexamcommittee | 李芳仁(Fang-Jen Lee),張鑫,鄧述諄(Shu-Chun Teng) | |
dc.subject.keyword | B型肝炎病毒,核心蛋白質,arginine-rich區域,細胞核定位訊號,細胞核輸出訊號,磷酸化, | zh_TW |
dc.subject.keyword | Hepatitis B virus,HBV,core protein,HBc,arginine-rich domain,ARD,nuclear localization signals,NLS,nuclear export signals,NES,phosphorylation, | en |
dc.relation.page | 54 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2010-08-18 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 分子醫學研究所 | zh_TW |
顯示於系所單位: | 分子醫學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-99-1.pdf 目前未授權公開取用 | 8.19 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。