[The effect of HCV NS5A protein on HCV IRES-dependent translation in HepG2 cells].
OBJECTIVE
To study the effect of HCV NS5A protein on HCV IRES-dependent translation in HepG2 cells.


METHODS
HepG2 cells were co-transfected with a plasmid vector containing a bicistronic transcript carrying Renilla luciferase and firefly luciferase genes separated by HCV IRES sequences, and an expressing vector producing the NS5A protein. The luciferase activity and the mRNA of the luciferase gene were then detected. The NS5A expression was confirmed by fluorescence microscopy.


RESULTS
HCV NS5A protein was detected in the cytoplasm of the HepG2 cells transfected with pcDNA-NS5A, and the luciferase activity was up-regulated in the presence of the HCV NS5A protein while the expression of luciferase mRNA showed no difference.


CONCLUSIONS
HCV NS5A protein can upregulate the HCV IRES activity and this effect is dose-dependent with NS5A.
Key laboratory of Infectious Diseases, Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.
Juan
J
Chen
Wei-xian
WX
Chen
Zhen-zhen
ZZ
Zhang
Ai-long
AL
Huang
D056945:Hep G2 Cells; D016174:Hepacivirus / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D010957:Plasmids; D014176:Protein Biosynthesis; D017433:Protein Structure, Secondary; D012270:Ribosomes / Q000378:metabolism*; D014162:Transfection; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D012324:RNA-Dependent RNA Polymerase
D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase