The NS5A protein of hepatitis C virus represses gene expression of hRPB10alpha, a common subunit of host RNA polymerases, through interferon regulatory factor-1 binding site.
The nonstructural (NS) 5A protein of hepatitis C virus (HCV) plays important roles in both viral RNA replication and modulation of the physiology of the host cell. Here we report that NS5A repressed gene expression of hRPB10alpha, a common subunit of host RNA polymerases (Pol), in hepatoma cell lines and Huh-7 cells harboring HCV replicon. Analysis of the hRPB10alpha promoter region revealed that interferon regulatory factor-1 binding element (IRF-E) was essential for its transcription. The IRF-E was responsible for the NS5A-mediated repression of the hRPB10alpha transcription and its induction by IRF-1 that is known to be induced by interferon-alpha. Electrophoretic mobility shift assay showed that IRF-1 bound to the IRF-E and the binding reduced when NS5A was expressed. NS5A appeared to negatively regulate IRF-1 expression, which might be partly responsible for the decrease of hRPB10alpha expression. NS5A expression moderately decreased promoter-independent Pol activity in vitro. Transcription of adenoviral genes that are dependent on Pol II or III and propagation of adenoviral genome were impaired in HeLa cells with stable NS5A expression. The results suggest that NS5A may partly modulate host cell transcription by the down-regulation of hRPB10alpha.
Jung
Cho-Rok
CR
Gene Therapy Research Unit, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Yusong, Daejeon 305-806, Republic of Korea.
Choi
Seeyoung
S
Im
Dong-Soo
DS
D000256:Adenoviridae / Q000235:genetics; D001665:Binding Sites; D002460:Cell Line; D012321:DNA-Directed RNA Polymerases / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D050856:Interferon Regulatory Factor-1 / Q000235:genetics / Q000378:metabolism*; D016898:Interferon-alpha / Q000378:metabolism; D014158:Transcription, Genetic; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*; D012324:RNA-Dependent RNA Polymerase
D012321:DNA-Directed RNA Polymerases; D050856:Interferon Regulatory Factor-1; D016898:Interferon-alpha; D017361:Viral Nonstructural Proteins; C494257:IRF1 protein, human; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase; C113275:RNA polymerase alpha subunit