Suppression of the ERK1/2 signaling pathway from HCV NS5A protein expressed by herpes simplex recombinant viruses.
Two herpes simplex type 1 (HSV-1) recombinant viruses carrying the hepatitis C virus (HCV) NS5A open reading frame under the control of the cytomegalovirus immediate early (IE) or a herpes simplex chimeric promoter (alpha 4 gamma 1UL19) were constructed and characterized. Expression studies showed that both HSV-NS5A recombinant viruses were able to express high levels of the NS5A protein in infected cells. Most importantly, using this system, we demonstrated that the NS5A protein interacts with the growth receptor-bound protein 2 (Grb2) and inhibits the phosphorylation of the extracellular signal-regulated kinases 1 and 2 (ERK1/2) in HeLa, NIH3T3 or liver infected cells. Thus, our studies confirm the ability of the NS5A protein to perturb the extracellular signal-regulated kinase (ERK) pathway in HeLa cells by the use of an alternative system for NS5A expression and extend this observation to additional cell lines. We conclude that HSV-based viral vectors may provide a useful system for studying the expression and selected functional properties of the HCV NS5A protein.
Georgopoulou
U
U
Molecular Virology Laboratory, Hellenic Pasteur Institute, Athens, Greece.
Caravokiri
K
K
Mavromara
P
P
D016475:3T3 Cells; D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D051380:GRB2 Adaptor Protein; D015967:Gene Expression Regulation, Viral; D006367:HeLa Cells; D016174:Hepacivirus / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D020935:MAP Kinase Signaling System; D051379:Mice; D020929:Mitogen-Activated Protein Kinase Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010766:Phosphorylation; D011506:Proteins / Q000378:metabolism; D014709:Vero Cells; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*; D012324:RNA-Dependent RNA Polymerase
D048868:Adaptor Proteins, Signal Transducing; D051380:GRB2 Adaptor Protein; D020929:Mitogen-Activated Protein Kinase Kinases; D011506:Proteins; D017361:Viral Nonstructural Proteins; C495948:GRB2 protein, human; C495949:Grb2 protein, mouse; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase