https://scholars.lib.ntu.edu.tw/handle/123456789/627780
標題: | Polypyrimidine-tract-binding protein is a component of the HCV RNA replication complex and necessary for RNA synthesis | 作者: | Aizaki, Hideki Choi, Keum S HELENE MINYI LIU Li, Yi-jia Lai, Michael M C |
關鍵字: | hepatitis C virus; PTB; RNA replication; lipid rafts; HEPATITIS-C VIRUS; INTERNAL RIBOSOMAL ENTRY; NUCLEAR RIBONUCLEOPROTEIN A1; MESSENGER-RNA; POLIOVIRUS RNA; INHIBITS TRANSLATION; SUBGENOMIC REPLICON; PICORNAVIRUS RNA; EXON DEFINITION; LA AUTOANTIGEN | 公開日期: | 七月-2006 | 出版社: | SPRINGER | 卷: | 13 | 期: | 4 | 起(迄)頁: | 469 | 來源出版物: | Journal of biomedical science | 摘要: | The machinery for hepatitis C virus (HCV) RNA replication is still poorly characterized. The relationship between HCV RNA replication and translation is also not clear. We have previously shown that a cellular protein polypyrimidine-tract-binding protein (PTB) binds to HCV RNA at several different sites and modulates HCV translation in several ways. Here we show that PTB also participates in RNA replication. By bromouridine triphosphate (BrUTP) labeling and confocal microscopy of cells harboring an HCV replicon, we showed that the newly synthesized HCV RNA was localized to distinct structures in the cytoplasm, which also contain PTB. Membrane flotation analysis demonstrated that a fraction of cytoplasmic PTB was associated with a detergent-resistant membrane (DRM) structure consisting of lipid rafts, which also contained HCV nonstructural proteins and the human vesicle-associated membrane protein-associated protein (hVAP-33). PTB in the DRM was resistant to protease digestion, but became sensitive after treatment with the raft-disrupting agents. PTB in the DRM consisted of multiple isoforms and the brain-specific paralog. By using small interfering RNA (siRNA) of PTB, we showed that silencing of the endogenous PTB reduced the replication of HCV RNA replicon. In a cell-free, de novo HCV RNA synthesis system, HCV RNA synthesis was inhibited by anti-PTB antibody. These studies together indicated that PTB is a part of the HCV RNA replication complex and participates in viral RNA synthesis. Thus, PTB has dual functions in HCV life cycle, including translation and RNA replication. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/627780 | ISSN: | 1021-7770 | DOI: | 10.1007/s11373-006-9088-4 |
顯示於: | 生物化學暨分子生物學科研究所 |
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