Newly synthesized hepatitis C virus replicon RNA is protected from nuclease activity by a protease-sensitive factor(s)
- PMID: 15331754
- PMCID: PMC514995
- DOI: 10.1128/JVI.78.18.10202-10205.2004
Newly synthesized hepatitis C virus replicon RNA is protected from nuclease activity by a protease-sensitive factor(s)
Abstract
Biochemical characterization of hepatitis C virus (HCV) replication using purified, membrane-associated replication complexes is hampered by the presence of endogenous nuclease activity that copurifies with the replication complex. In this study, pulse-chase analyses were used to demonstrate that newly synthesized replicon RNA was protected from nuclease activity by a factor(s) that was sensitive to 0.5% NP-40 or protease treatment. Nuclease susceptibility was not related to disruption of lipid membranes, since NP-40 did not significantly affect the buoyant density of HCV replication complexes or protease susceptibility of HCV NS3 and NS5A proteins. These results suggest that a protease-sensitive factor(s) protects newly synthesized RNA from nuclease degradation.
Copyright 2004 American Society for Microbiology
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