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Figure 9 | Virology Journal

Figure 9

From: Involvement of PKR and RNase L in translational control and induction of apoptosis after Hepatitis C polyprotein expression from a Vaccinia virus recombinant

Figure 9

Expression of HCV polyprotein from VV induces ribosomal RNA degradation mediated by RNaseL and triggers apoptosis through RNase L independently of PKR. A: Monolayers of HeLa cells were either uninfected (U), single-infected with VT7 (5 PFU/cell), single-infected with VT7-HCV7.9 (5 PFU/cell) in the presence (+) or absence (-) of IPTG, or triple-infected with VV-RL + VT7 + VV-25AS (2 PFU of each virus/cell) (C+). Infections proceeded for 24 hours. 2 μg of total RNA was fractionated in 1% agarose-formaldehyde gel and stained with ethidium bromide. Abundant ribosomal RNAs 28S and 18S are indicated. B and C: PKR knockout (PKR-/-) and PKR WT cells (PKR+/+) (panel B), as well as RNase L knockout (RL-/-) and RNase L WT cells (RL+/+) (panel C), were infected at 5 PFU/cell with the recombinant VT7-HCV7.9 virus, in the presence (+) or absence (-) of the inductor IPTG. The apoptotic levels in cell extracts were determined at 24 h.p.i. by ELISA. The recombinant VV-PKR virus was used as a control. U: Uninfected cells.

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