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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張鑫 | zh_TW |
| dc.contributor.author | 戴于晴 | zh_TW |
| dc.contributor.author | Yu-Ching Dai | en |
| dc.date.accessioned | 2021-05-19T17:48:17Z | - |
| dc.date.available | 2023-12-13 | - |
| dc.date.copyright | 2018-03-29 | - |
| dc.date.issued | 2018 | - |
| dc.date.submitted | 2002-01-01 | - |
| dc.identifier.citation | 1. Feinstone SM, et al., Transfusion-associated hepatitis not due to viral hepatitis type A or B. N Engl J Med. 1975 Apr 10;292(15):767-70.
2. Choo QL, et al., Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science. 1989 Apr 21;244(4902):359-62. 3. Darius Moradpour, François Penin and Charles M. Rice, Replication of hepatitis C virus. Nature Reviews Microbiology 5, 453-463 (June 2007) 4. Nakano T, et al., An updated analysis of hepatitis C virus genotypes and subtypes based on the complete coding region. Liver Int. 2011, December 32 (2): 339–45. 5. Wilkins T, et al., Hepatitis C: diagnosis and treatment. Am Fam Physician. 2010 Jun 1;81(11):1351-7. 6. Paul D, Madan V and Bartenschlager R, Hepatitis C virus RNA replication and assembly: living on the fat of the land. Cell Host Microbe. 2014 Nov 12;16 (5):569-79. 7. Ashfaq UA, et al., An overview of HCV molecular biology, replication and immune responses. Virol J. 2011 Apr 11;8:161. 8. Anzola M and Burgos JJ, Hepatocellular carcinoma: molecular interactions between hepatitis C virus and p53 in hepatocarcinogenesis. Expert Rev Mol Med. 2003 Nov 19;5(28):1-16. 9. Scheel TK and Rice CM. Understanding the hepatitis C virus life cycle paves the way for highly effective therapies. Nat Med. 2013 Jul;19(7):837-49. 10. Kohichiroh Yasui, et al., The Native Form and Maturation Process of Hepatitis C Virus Core Protein. J Virol. 1998 Jul; 72(7): 6048–6055. 11. Chang SC, et al., Nuclear localization signals in the core protein of hepatitis C virus. Biochem Biophys Res Commun. 1994 Dec 15;205(2):1284-90. 12. Ryosuke Suzuki, et al., Molecular Determinants for Subcellular Localization of Hepatitis C Virus Core Protein. J Virol. 2005 Jan; 79(2): 1271–1281. 13. Herker E, et al., Efficient hepatitis C virus particle formation requires diacylglycerol acyltransferase-1. Nat Med. 2010 Nov;16(11):1295-8. 14. Menzel N, et al., MAP-kinase regulated cytosolic phospholipase A2 activity is essential for production of infectious hepatitis C virus particles. PLoS Pathog. 2012;8(7):e1002829. 15. Neveu G, et al., Identification and targeting of an interaction between a tyrosine motif within hepatitis C virus core protein and AP2M1 essential for viral assembly. PLoS Pathog. 2012;8(8):e1002845. 16. Takahiro Masaki, et al., Interaction of Hepatitis C Virus Nonstructural Protein 5A with Core Protein Is Critical for the Production of Infectious Virus Particles. J Virol. 2008 Aug; 82(16): 7964–7976. 17. V Deleersnyder, et al., Formation of native hepatitis C virus glycoprotein complexes. J Virol. 1997 Jan; 71(1): 697–704. 18. Bartosch B, et al., Cell entry of hepatitis C virus requires a set of co-receptors that include the CD81 tetraspanin and the SR-B1 scavenger receptor. J Biol Chem. 2003 Oct 24;278(43):41624-30. 19. Steinmann E, et al., Hepatitis C virus p7 protein is crucial for assembly and release of infectious virions. PLoS Pathog. 2007 Jul;3(7):e103. 20. Reed KE, Grakoui A and Rice CM, Hepatitis C virus-encoded NS2-3 protease: cleavage-site mutagenesis and requirements for bimolecular cleavage. J Virol. 1995 Jul;69(7):4127-36. 21. Y. Yan, et al., Complex of NS3 protease and NS4A peptide of BK strain hepatitis C virus: a 2.2 A resolution structure in a hexagonal crystal form. Protein Sci. 1998 Apr; 7(4): 837–847. 22. Pang PS, Jankowsky E, Planet PJ and Pyle AM. The hepatitis C viral NS3 protein is a processive DNA helicase with cofactor enhanced RNA unwinding. EMBO J. 2002 Mar 1;21(5):1168-76. 23. Denise Egger, et al., Expression of Hepatitis C Virus Proteins Induces Distinct Membrane Alterations Including a Candidate Viral Replication Complex. J Virol. 2002 Jun; 76(12): 5974–5984. 24. Evans MJ, Rice CM and Goff SP, Phosphorylation of hepatitis C virus nonstructural protein 5A modulates its protein interactions and viral RNA replication. Proc Natl Acad Sci U S A. 2004 Aug 31;101(35):13038-43. 25. André P, et al., Characterization of low- and very-low-density hepatitis C virus RNA-containing particles. J Virol. 2002 Jul;76(14):6919-28. 26. Suzuki T, Morphogenesis of infectious hepatitis C virus particles. Front Microbiol. 2012 Feb 7;3:38. 27. Scheel TK and Rice CM, Understanding the hepatitis C virus life cycle paves the way for highly effective therapies. Nat Med. 2013 Jul;19(7):837-49. 28. Lindenbach BD and Rice CM, The ins and outs of hepatitis C virus entry and assembly. Nat Rev Microbiol. 2013 Oct;11(10):688-700. 29. Gabrielle Vieyres, Jean Dubuisson, and Thomas Pietschmann, Incorporation of Hepatitis C Virus E1 and E2 Glycoproteins: The Keystones on a Peculiar Virion. Viruses 2014, 6(3), 1149-1187. 30. Lupberger J, et al., EGFR and EphA2 are host factors for hepatitis C virus entry and possible targets for antiviral therapy. Nat Med. 2011;17:589–595. 31. Sainz B, et al., Identification of the Niemann-Pick C1-like 1 cholesterol absorption receptor as a new hepatitis C virus entry factor. Nat Med. 2012;18:281–285. 32. Owen DM, Huang H, Ye J, and Gale M., Apolipoprotein E on hepatitis C virion facilitates infection through interaction with low-density lipoprotein receptor. J. Virol. 2009;394:99–108. 33. Blanchard E, et al., Hepatitis C virus entry depends on clathrin-mediated endocytosis. J Virol 2006; 80: 6964-6972 34. Harris HJ, et al., CD81 and claudin 1 coreceptor association: role in hepatitis C virus entry. J Virol. 2008;82:5007–5020. 35. Harris HJ, et al., Claudin association with CD81 defines hepatitis C virus entry. J Biol Chem. 2010;285:21092–21102. 36. Farquhar MJ, et al., Hepatitis C virus induces CD81 and claudin-1 endocytosis. J Virol. 2012;86:4305–4316. 37. Michael P. Manns, et al., Hepatitis C virus infection. Nat Rev Dis Primers. 2017 Mar 2;3:17006. 38. Alvisi G, Madan V and Bartenschlager R, Hepatitis C virus and host cell lipids: an intimate connection. RNA Biol. 2011;8:258–269. 39. Tscherne DM, et al., Time- and temperature-dependent activation of hepatitis C virus for low-pH-triggered entry. J Virol. 2006;80:1734–1741. 40. Gosert R, et al., Identification of the hepatitis C virus RNA replication complex in Huh-7 cells harboring subgenomic replicons. J Virol. 2003;77:5487–5492. 41. Bartenschlager R, Cosset FL and Lohmann V, Hepatitis C virus replication cycle. J Hepatol. 2010;53:583–585. 42. Miyanari, Y. et al. Hepatitis C virus non-structural proteins in the probable membranous compartment function in viral genome replication. J. Biol. Chem., 2003, 278, 50301–50308. 43. McLauchlan J, Lipid droplets and hepatitis C virus infection. Biochim Biophys Acta 2009, 1791:552-559. 44. Filipe A, McLauchlan J, Hepatitis C virus and lipid droplets: finding a niche. Trends Mol Med 2015, 21:34-42. 45. Huang H, et al., Hepatitis C virus production by human hepatocytes dependent on assembly and secretion of very low-density lipoproteins. Proc Natl Acad Sci U S A. 2007 Apr 3; 104(14):5848-53 46. Gastaminza P, et al., Cellular determinants of hepatitis C virus assembly, maturation, degradation, and secretion. J Virol. 2008 Mar; 82(5):2120-9. 47. Wang YH, et al., Novel nuclear export signal-interacting protein, NESI, critical for the assembly of hepatitis delta virus. J Virol. 2005 Jul; 79(13):8113-20 48. Rutsch F, et al., Identification of a putative lysosomal cobalamin exporter altered in the cblF defect of vitamin B12 metabolism. Nat Genet. 2009 Feb; 41(2):234-9. 49. Coelho D, et al., Mutations in ABCD4 cause a new inborn error of vitamin B12 metabolism. Nat Genet. 2012; 44: 1152–5. 50. Linda Tzu-Ling, et al., LMBD1 Protein Serves as a Specific Adaptor for Insulin Receptor Internalization. J Biol Chem. 2013 Nov 8; 288(45): 32424–32432. 51. Maria Victoria Preciado, et al., Hepatitis C virus molecular evolution: Transmission, disease progression and antiviral therapy. World J Gastroenterol, 2014, November 21; 20(43): 15992-16013 52. Yen-Ju Chen, 2016, Cellular protein LMBD1 regulates RNA replication of Hepatitis C virus. Graduate Institute of Microbiology College of Medicine, National Taiwan University 53. Kosuke Kawaguchi, et al., Translocation of the ABC transporter ABCD4 from the endoplasmic reticulum to lysosomes requires the escort protein LMBD1. Sci Rep. 2016; 6: 30183. 54. Chun-Wei Chang, 2012, LMBD1 protein involves in multiplication of hepatitis C virus. Graduate Institute of Biochemistry and Molecular Biology College of Medicine, National Taiwan University 55. Blight KJ, McKeating JA and Rice CM, Highly permissive cell lines for subgenomic and genomic hepatitis C virus RNA replication. J Virol. 2002 Dec;76(24):13001-14. 56. Wu SC, et al., Hepatitis C virus NS5A protein down-regulates the expression of spindle gene Aspm through PKR-p38 signaling pathway. J Biol Chem. 2008 Oct 24;283(43):29396-404. 57. Chih-Hsien Kao, 2008, Selection of host factors and chemicals that regulate the replication of hepatitis C virus by using a newly established infectious clone. Graduate Institute of Biochemistry and Molecular Biology College of Medicine, National Taiwan University 58. Ya-Li Tsai, 2002, Establishment of a recombinant adenovirus-inducible system to express the hepatitis C virus core protein. Graduate Institute of Biochemistry and Molecular Biology College of Medicine, National Taiwan University 59. Ping-Kun Hsieh, et al., Assembly of Severe Acute Respiratory Syndrome Coronavirus RNA Packaging Signal into Virus-Like Particles Is Nucleocapsid Dependent. J Virol. 2005 Nov; 79(22): 13848–13855. 60. Gary J. Doherty and Harvey T. McMahon, Mechanisms of Endocytosis. Annu. Rev. Biochem. 2009. 78:857–902 61. Laurent Meertens, Claire Bertaux, and Tatjana Dragic, Hepatitis C Virus Entry Requires a Critical Postinternalization Step and Delivery to Early Endosomes via Clathrin-Coated Vesicles. J Virol. 2006 Dec; 80(23): 11571–11578. 62. Tosoni D, et al., TTP specifically regulates the internalization of the transferrin receptor. Cell. 2005 Dec 2;123(5):875-88. 63. Nathan L Meyers, et al., Entangled in a membranous web: ER and lipid droplet reorganization during hepatitis C virus infection. Current Opinion in Cell Biology 2016, 41:117–124 64. Kang SM, et al., Regulation of hepatitis C virus replication by the core protein through its interaction with viral RNA polymerase. Biochem Biophys Res Commun. 2009 Aug 14;386(1):55-9. 65. Ikeda M1, Yi M, Li K and Lemon SM, Selectable subgenomic and genome-length dicistronic RNAs derived from an infectious molecular clone of the HCV-N strain of hepatitis C virus replicate efficiently in cultured Huh7 cells. J Virol. 2002 Mar;76(6):2997-3006. 66. NIZAR N. ZEIN, Clinical Significance of Hepatitis C Virus Genotypes. Clin Microbiol Rev. 2000 Apr;13(2):223-35. 67. Mar Rodríguez-Rodrígueza, et al., Fusogenic properties of the Ectodomain of HCV E2 envelope protein. BBA - Biomembranes 1860 (2018) 728–736 | - |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/7627 | - |
| dc.description.abstract | C型肝炎病毒 (hepatitis C virus, HCV) 主要經由血液和體液傳染，感染後會造成C型肝炎，屬於全球性的感染疾病。HCV具有正向單股RNA基因體，在感染宿主細胞後,會以其正向單股RNA基因體進行轉譯，並在內質網的周邊形成適合病毒複製的特殊網狀結構，藉由細胞中油滴的幫助，進行病毒顆粒的組裝。本實驗室先前的研究發現，在細胞中表現LMBD1重組蛋白質時，此蛋白質與細胞中的油滴有明顯共位的現象。此外，在細胞中同時送入LMBD1以及HCV core蛋白質的表現質體後，也可觀察到LMBD1和core蛋白質的共位現象。利用共同免疫沉澱分析發現LMBD1以及core兩蛋白質在細胞中有交互作用。另外，在踢弱 (knockdown) LMBRD1基因表現的細胞感染HCV後，其細胞中的HCV負向基因體RNA量有上升的趨勢。為了更深入的瞭解LMBRD1基因與HCV基因體複製與病毒增殖的相關性，本研究利用HCVR次基因體細胞以及在HCV細胞培養系統中所產生的病毒 (HCVcc) 進行研究。結果顯示，在HCVR細胞中LMBRD1基因表現缺失會造成HCV的RNA負向基因體顯著降低，而其基因產物LMBD1以及NESI對於HCV基因體RNA複製皆有正向的調控作用。此外，HCV core蛋白質表現在踢弱LMBRD1基因的細胞中會使得HCV負向基因體量增加，推測core蛋白質可能會透過LMBD1蛋白質的幫助，穩定基因體RNA複製推進至病毒包裹的動態平衡。另一方面，在踢弱LMBRD1基因表現的情況下，被HCVcc感染的細胞所產生出來的病毒顆粒數量並無明顯差異，但其感染能力明顯降低。LMBRD1基因的表現影響了HCV基因體的複製以及病毒顆粒的組裝，然而其中的機轉仍然有待進一步的研究。 | zh_TW |
| dc.description.abstract | Hepatitis C virus (HCV) is mainly transmitted by blood and body fluids. It causes hepatitis C after infection, which is a worldwide infectious disease. HCV possesses positive single-strand RNA genome. Upon infection with a host cell, the positive single-strand RNA acts as a template and translates to viral proteins. In the HCV-infected cells, HCV induces formation of a special membrane structure around the endoplasmic reticulum suitable for the replication of viral genome. In addition, assembly of HCV particles is helped by lipid droplets. Previous studies from our laboratory demonstrated that the LMBD1 recombinant proteins colocalized with the lipid droplet marker ADRP in the cells. Co-localization of LMBD1 with the viral core protein was also detected when co-expression of LMBD1 and HCV core protein. Co-immunoprecipitation analysis revealed that LMBD1 protein interacts with core protein. In addition, the level of HCV antigenomic RNA tends to be higher in HCVcc-infected Huh7.5 cells to which LMBRD1 gene has been knocked-down (Huh-shLMBRD1) compared to the control Huh7.5 cells to which LUC gene has been knocked-down (Huh-shLUC). In this study, HCVR subgenomic replicon cells and HCVcc infection system were applied to further examine the roles of LMBRD1 gene on HCV replication and assembly. The results show a significant lower level of HCV antigenomic RNA in LMBRD1-knockdown HCVR cells compared to the control LUC-knockdown cells, both LMBRD1 gene products LMBD1 and NESI had positive effects on HCV replication. In addition, HCV core protein expression in LMBRD1-knockdown HCVR cells increased the levels of HCV antigenomic RNA, suggesting that core protein may interact with LMBD1 to stablize the turn over between viral replication and assembly. On the other hand, HCVcc produced from HCVcc-infected Huh-shLMBRD1 cells (HCVcc-shLMBRD1) had no significant difference in the number of virus particles compared to the control HCVcc produced from HCVcc-infected Huh-shLUC cells (HCVcc-shLUC) but significantly reduced the infectivity. Expression of LMBRD1 gene affects HCV replication and assembly, the detailed mechanism remains to be further elucidated. | en |
| dc.description.provenance | Made available in DSpace on 2021-05-19T17:48:17Z (GMT). No. of bitstreams: 1 ntu-107-R04445119-1.pdf: 1822573 bytes, checksum: b1f7c39923ce46d57be9021a2088b5d5 (MD5) Previous issue date: 2018 | en |
| dc.description.tableofcontents | 中文摘要 i
Abstract ii Contents iv Content of figures vii I. Introduction 1 A. Hepatitis C virus (HCV) 1 a. History 1 b. Virology 1 c. The structure and genome of HCV 2 d. Life cycle of HCV 6 i.Attachment and entry 6 ii.Replication 7 iii.Assembly and release 8 B. Limb region 1 (LMBR1) domain containing 1 gene (LMBRD1) 8 II. Specific aims 10 III. Materials and Methods 12 A. Materials 12 a. Chemicals and reagents 12 b. Enzymes 13 c. Antibodies 14 d. Kits 14 e. Cell lines 15 f. Plasmids 16 B. Methods 18 a. Construction of plasmid pLenti-C-NESI-shR-mGFP 18 b. Transfection 18 c. Lentiviral Transduction 19 d. Western blot 19 e. Quantification of viral RNA and reverse transcription-qPCR (RT-qPCR) 20 f. HCV cell culture system (HCVcc system) 21 g. Harvest and purification of HCVcc 21 IV. Results 23 A. Knocking down LMBRD1 gene reduced HCV replication. 23 B. Both LMBD1 and NESI expression in LMBRD1-knockdown cell rescue HCV replication. 23 C. LMBRD1 gene expression might disrupt HCV replication in HCVcc-infected Huh7.5 cells. 24 D. Core expression in LMBRD1 knockdown cell increases the levels of HCV antigenomic RNA. 24 E. LMBRD1 gene expression may contribute to the production of infectious HCVcc. 25 F. LMBRD1 gene might play a crucial role in HCVcc propagation. 26 G. LMBRD1 gene knockdown might change the properties of HCVcc resulting in a reduced ability in cell-entry. 26 H. The HCVcc-shLMBRD1 has poor replication and protein expression in naïve Huh7.5 cells. 27 V. Discussion 28 A. LMBRD1 gene knockdown inhibits HCV replication in HCVR cells. 29 B. LMBRD1 gene knockdown may have impact on the effect of HCV core protein in HCV replication. 30 C. LMBRD1 gene expression may have no effect on viral release but impact on viral assembly, leading to a poor infectivity. 31 D. The replication of HCVcc-shLMBRD1 in cells is significantly less efficient. 32 VI. Figures 34 VII. References 46 VIII. Appendix Figures 53 | - |
| dc.language.iso | en | - |
| dc.subject | Ｃ型肝炎病毒 | zh_TW |
| dc.subject | 病毒組裝 | zh_TW |
| dc.subject | 病毒基因體複製 | zh_TW |
| dc.subject | LMBD1蛋白質 | zh_TW |
| dc.subject | LMBRD1基因 | zh_TW |
| dc.subject | Ｃ型肝炎病毒 | zh_TW |
| dc.subject | LMBRD1基因 | zh_TW |
| dc.subject | LMBD1蛋白質 | zh_TW |
| dc.subject | 病毒基因體複製 | zh_TW |
| dc.subject | 病毒組裝 | zh_TW |
| dc.subject | LMBD1 protein | en |
| dc.subject | viral assembly | en |
| dc.subject | viral genome replication | en |
| dc.subject | LMBD1 protein | en |
| dc.subject | LMBRD1 gene | en |
| dc.subject | Hepatitis C virus | en |
| dc.subject | viral assembly | en |
| dc.subject | viral genome replication | en |
| dc.subject | Hepatitis C virus | en |
| dc.subject | LMBRD1 gene | en |
| dc.title | LMBRD1基因產物在C型肝炎病毒RNA複製及病毒組裝中扮演的角色 | zh_TW |
| dc.title | Roles of LMBRD1 gene products involved in the replication and assembly of hepatitis C virus | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 106-1 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 陳美如;楊宏志 | zh_TW |
| dc.contributor.oralexamcommittee | ;; | en |
| dc.subject.keyword | Ｃ型肝炎病毒,LMBRD1基因,LMBD1蛋白質,病毒基因體複製,病毒組裝, | zh_TW |
| dc.subject.keyword | Hepatitis C virus,LMBRD1 gene,LMBD1 protein,viral genome replication,viral assembly, | en |
| dc.relation.page | 54 | - |
| dc.identifier.doi | 10.6342/NTU201800389 | - |
| dc.rights.note | 同意授權(全球公開) | - |
| dc.date.accepted | 2018-02-09 | - |
| dc.contributor.author-college | 醫學院 | - |
| dc.contributor.author-dept | 微生物學研究所 | - |
| 顯示於系所單位： | 微生物學科所 | |
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