親和力純化與蛋白質體學研究顯示hnRNPH1、NF45和C14orf166與C型肝炎病毒蛋白鞘有交互作用並影響病毒複製及寄主細胞的增生

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親和力純化與蛋白質體學研究顯示hnRNPH1、NF45和C14orf166與C型肝炎病毒蛋白鞘有交互作用並影響病毒複製及寄主細胞的增生

An affinity purification-based proteomic approach reveals hnRNPH1, NF45, and C14orf166 as three HCV core-interacting proteins involved in viral replication and cell proliferation in host cells

李峻瑋 , Ph.D　　Advisor：王憲威　　

英文

C型肝炎病毒；核心蛋白；液相層析串聯式質譜儀；異質核醣核酸蛋白H1 ；核因子； C14orf166 ；增殖；病毒複製； HCV ； HCVc ； LC-MS/MS ； hnRNPH1 ； NF45 ； C14orf166 ； proliferation ； viral replication

Abstract │ Reference (82) │ Altmetrics

Abstract 〈TOP〉

C型肝炎病毒核心蛋白（HCVc）構成病毒核酸蛋白鞘，並可能透過與宿主細胞因子互動以調控其病毒複製及本身之細胞功能，從而導致C型肝炎病毒持續感染及發病。HCVc174是1b基因型HCV（HCV-1b）的成熟型HCVc。利用了N端生物素或C端鈣調蛋白結合肽／蛋白質A標籤的HCVc174親和性純化與一維電泳和串連質譜儀技術（LC-MS/MS），找到了36個極有可能HCVc174與有交互作用的細胞蛋白。藉由沉澱法與共軛焦成像技術，進一步確認異質核醣核酸蛋白H1（hnRNPH1）、核因子（NF45）及C14orf166是新穎的成熟型HCVc174結合寄主蛋白。此外，NF45是透過RNA機制與HCVc174結合。這三種細胞蛋白質在細胞質和細胞核中皆與異位表達的HCVc-1b共位而存，顯示這三種細胞蛋白質在HCVc生成後與自然移位的HCVc174產生空間上的相互作用。然而此共存現象在1b或2a基因型之病毒感染複製時，卻大部分轉移到細胞質，說明活躍的病毒複製會將這些有交互作用的蛋白質侷限在細胞質中。上述結果顯示，hnRNPH1、NF45和C14orf166與HCVc174的空間相互作用，或許可以在急慢性C型肝炎病毒感染期間，發揮控制HCV複製或影響細胞功能的作用。進一步確認了這三種細胞蛋白在細胞存活與C型肝炎病毒複製方面的功能相關性，以查明這三種細胞蛋白成為蛋白質性抗病毒藥物的潛力。透過使之個別降低或過度表達，發現這三種細胞基因是肝細胞長期生存的必要條件，同時也是抑制HCV複製程序的控制因子。本研究中選用C14orf166為對象，進一步研究其與HCVc在HCV複製過程中的交互作用關係。研究發現無論HCVc是否以反式（in trans）存在，C14orf166的過度表達皆可減弱HCV複製；而不論順式（in cis）或反式（in trans）的HCVc表達對於受C14orf166控制的HCV複製程序均無任何顯著的協同或對抗效應。至於HCVc是否僅在C14orf166調控下的HCV複製程序中扮演旁觀輔助因子的角色，或是在生物程序的其他步驟中扮演積極參與因子的角色，則有待進一步研究釐清。本研究結果顯示hnRNPH1、NF45與C14orf166為細胞生存的必要條件，且對於HCV複製過程展現負向控制作用。因此，這三種基因的過度表達可能有助於抑制HCV複製。或者，可針對這三種與HCVc互動的細胞蛋白質，以其功能區段基因產物做為今後研發胜肽型抗病毒藥物的基礎。

Parallel Abstract 〈TOP〉

The hepatitis C virus core protein (HCVc) forms the viral nucleocapsid and participates in viral persistence and pathogenesis, possibly by interacting with host cellular factors to modulate viral replication and cellular functions. HCVc174 is a mature form of HCVc from HCV genotype 1b (HCV-1b). I identified 36 cellular protein candidates using one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and liquid chromatography–mass spectrometry/mass spectrometry-based proteomics after affinity purification with HCVc174 that was tagged N-terminally with biotin or C-terminally with calmodulin-binding peptide/protein A. Using pull-down and confocal imaging techniques, I confirmed that heterogeneous nuclear ribonucleoprotein H1 (hnRNPH1), nuclear factor 45 (NF45), and chromosome 14 open reading frame 166 (C14orf166) are novel HCVc174-interacting host proteins. In addition, NF45 interacts with HCVc174 in an RNA-dependent manner. The 3 cellular proteins colocalized with ectopic HCVc-1b in the cytoplasm and nucleus, which indicates that they interact spatially with naturally translocated HCVc174 after HCVc biogenesis. Such colocalization, however, shifted to the cytoplasm with the replicating virus of the 1b or 2a genotype, which suggests that active viral replication confines these interacting proteins to the cytoplasm. These results demonstrate that spatial interactions of hnRNPH1, NF45, and C14orf166 with HCVc174 likely modulate HCV replication or cellular function during acute and chronic HCV infection. To investigate the potential of these 3 cellular proteins as protein-based antiviral drugs, their functional relevance in HCV replication and cell survival was preliminarily determined. Through individual silencing or overexpression, it was found that the 3 cellular genes are required for the prolonged survival of hepatocytes and are negative regulators of the active HCV replication process. C14orf166 was selected to further study the interactive relationship of HCVc in HCV replication. Overexpression of C14orf166 attenuated HCV subgenome replication regardless of whether HCVc was present in trans, and HCVc expression in cis or in trans did not have any significant synergistic or counter effects on C14orf166-modulated HCV replication. Whether HCVc is merely a bystander co-factor participating in the C14orf166-mediated HCV replication process or an active co-factor in other steps of the biological process requires further investigation. Our results show that hnRNPH1, NF45, and C14orf166 are required for cell survival and can negatively regulate HCV replication. Consequently, the overexpression of these 3 genes may be beneficial in limiting HCV replication. Alternatively, truncated gene products of these 3 cellular proteins encoding domains that interact with the HCV core may be useful as potential peptide-based antiviral drugs.

Reference ( 82 ) 〈TOP〉

1. Blackard, J. T.; Kemmer, N.; Sherman, K. E., Extrahepatic replication of HCV: insights into clinical manifestations and biological consequences. Hepatology 2006, 44, (1), 15-22.
連結：
2. Sansonno, D.; Dammacco, F., Hepatitis C virus, cryoglobulinaemia, and vasculitis: immune complex relations. Lancet Infect Dis 2005, 5, (4), 227-36.
連結：
3. Ai, L. S.; Lee, Y. W.; Chen, S. S. L., Characterization of hepatitis C virus core protein multimerization and membrane envelopment: revelation of a cascade of core-membrane interactions. J Virol 2009, 83, (19), 9923-39.
連結：
4. Boonstra, A.; van der Laan, L. J.; Vanwolleghem, T.; Janssen, H. L., Experimental models for hepatitis C viral infection. Hepatology 2009, 50, (5), 1646-55.
連結：
5. Boni, S.; Lavergne, J. P.; Boulant, S.; Cahour, A., Hepatitis C virus core protein acts as a trans-modulating factor on internal translation initiation of the viral RNA. J Biol Chem 2005, 280, (18), 17737-48.
連結：

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