Ethanol facilitates hepatitis C virus replication via up-regulation of GW182 and heat shock protein 90 in human hepatoma cells.
Alcohol use and hepatitis C virus (HCV) infection synergize to cause liver damage, and microRNA-122 (miR-122) appears to play a key role in this process. Argonaute 2 (Ago2), a key component of the RNA-induced silencing complex (RISC), has been shown to be important in modulating miR-122 function during HCV infection. However, GW182, a critical component of processing bodies (GW bodies) that is recruited by Ago2 to target messenger RNA (mRNA), has not been assessed in HCV infection. To characterize the role of GW182 in the pathogenesis of HCV infection, we determined its transcription and protein expression in an HCV J6/JFH1 culture system. Transcript and protein levels of GW182 as well as HCV RNA and protein expression increased with alcohol exposure. Specific silencing of mRNA expression by small interfering RNA against GW182 significantly decreased HCV RNA and protein expression. Overexpression of GW182 significantly increased HCV RNA and protein expression in HCV J6/JFH1 infected Huh7.5 cells. Furthermore, GW182 colocalized and coimmunoprecipitated with heat shock protein 90 (HSP90), which increased upon alcohol exposure with and without HCV infection and enhanced HCV gene expression. The use of an HSP90 inhibitor or knockdown of HSP90 decreased GW182 and miR-122 expression and significantly reduced HCV replication.


CONCLUSIONS
Overall, our results suggest that GW182 protein that is linked to miR-122 biogenesis and HSP90, which has been shown to stabilize the RISC, are novel host proteins that regulate HCV infection during alcohol abuse.
Department of Medicine, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
Terence N
TN
Bukong
Wei
W
Hou
Karen
K
Kodys
Gyongyi
G
Szabo
D000891:Anti-Infective Agents, Local / Q000494:pharmacology*; D001324:Autoantigens / Q000378:metabolism*; D045744:Cell Line, Tumor; D000431:Ethanol / Q000494:pharmacology*; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D016174:Hepacivirus / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism*; D015854:Up-Regulation; D014764:Viral Proteins / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects*
D000891:Anti-Infective Agents, Local; D001324:Autoantigens; D018841:HSP90 Heat-Shock Proteins; C531907:MIRN122 microRNA, human; D035683:MicroRNAs; D016601:RNA-Binding Proteins; C465663:TNRC6A protein, human; D014764:Viral Proteins; D000431:Ethanol