Abstract:
:Chronic infection by hepatitis C virus (HCV) can lead to severe hepatitis and cirrhosis and is closely associated with hepatocellular carcinoma. The replication cycle of HCV is poorly understood but is likely to involve interaction with host factors. In this report, we show that NS5B, the HCV RNA-dependent RNA polymerase (RdRp), interacts with a human RNA helicase, p68. Transient expression of NS5B alone, as well as the stable expression of all the nonstructural proteins in a HCV replicon-bearing cell line (V. Lohmann, F. Korner, J.-O. Koch, U. Herian, L. Theilmann, and R. Bartenschlager, Science 285:110-113), causes the redistribution of endogenous p68 from the nucleus to the cytoplasm. Deletion of the C-terminal two-thirds of NS5B (NS5BDeltaC) dramatically reduces its coimmunoprecipitation (co-IP) with endogenous p68, while the deletion of the N-terminal region (NS5BDeltaN1 and NS5BDeltaN2) does not affect its interaction with p68. In consistency with the co-IP results, NS5BDeltaC does not cause the relocalization of p68 whereas NS5BDeltaN1 does. With a replicon cell line, we were not able to detect a change in positive- and negative-strand synthesis when p68 levels were reduced using small interfering RNA (siRNA). In cells transiently transfected with a full-length HCV construct, however, the depletion (using specific p68 siRNA) of endogenous p68 correlated with a reduction in the transcription of negative-strand from positive-strand HCV RNA. Overexpression of NS5B and NS5BDeltaN1, but not that of NS5BDeltaC, causes a reduction in the negative-strand synthesis, indicating that overexpressed NS5B and NS5BDeltaN1 sequesters p68 from the replication complexes (thus reducing their replication activity levels). Identification of p68 as a cellular factor involved in HCV replication, at least for cells transiently transfected with a HCV expression construct, is a step towards understanding HCV replication.
journal_name
J Viroljournal_title
Journal of virologyauthors
Goh PY,Tan YJ,Lim SP,Tan YH,Lim SG,Fuller-Pace F,Hong Wdoi
10.1128/jvi.78.10.5288-5298.2004subject
Has Abstractpub_date
2004-05-01 00:00:00pages
5288-98issue
10eissn
0022-538Xissn
1098-5514journal_volume
78pub_type
杂志文章abstract::Diminished in vitro proliferation of human immunodeficiency virus type 1 (HIV-1)-specific CD4+T cells has been associated with HIV-1 viremia and declining CD4+ T-cell counts during chronic infection. To better understand this phenomenon, we examined whether HIV-1 Gag p24 antigen-induced CD4+ T-cell proliferation might...
journal_title:Journal of virology
pub_type: 杂志文章
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doi:10.1128/JVI.05756-11
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doi:10.1128/JVI.27.3.677-687.1978
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pub_type: 杂志文章
doi:10.1128/JVI.68.7.4251-4261.1994
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pub_type: 杂志文章
doi:10.1128/JVI.7.6.707-719.1971
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pub_type: 杂志文章
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更新日期:1983-01-01 00:00:00
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pub_type: 杂志文章
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journal_title:Journal of virology
pub_type: 杂志文章
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pub_type: 杂志文章
doi:10.1128/JVI.26.2.380-388.1978
更新日期:1978-05-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.64.12.5919-5926.1990
更新日期:1990-12-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/JVI.02514-09
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pub_type: 杂志文章
doi:10.1128/JVI.73.7.5698-5706.1999
更新日期:1999-07-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.69.12.7791-7799.1995
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pub_type: 杂志文章
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pub_type: 杂志文章
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更新日期:1970-07-01 00:00:00
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journal_title:Journal of virology
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.63.2.696-704.1989
更新日期:1989-02-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
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更新日期:2006-02-01 00:00:00
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pub_type: 杂志文章
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更新日期:1979-12-01 00:00:00