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Journal of Virology, September 2004, p. 10202-10205, Vol. 78, No. 18
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.18.10202-10205.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Newly Synthesized Hepatitis C Virus Replicon RNA Is Protected from Nuclease Activity by a Protease-Sensitive Factor(s)

Guang Yang, Daniel C. Pevear, Marc S. Collett, Srinivas Chunduru, Dorothy C. Young, Christopher Benetatos, and Robert Jordan*

ViroPharma Incorporated, Exton, Pennsylvania

Received 4 March 2004/ Accepted 30 April 2004

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.

* Corresponding author. Present address: SIGA Technologies, Inc., 4575 S.W. Research Way, Corvallis, OR 97333. Phone: (541) 753-2000. Fax: (541) 753-9999. E-mail: rjordan{at}sgph.com.

Journal of Virology, September 2004, p. 10202-10205, Vol. 78, No. 18
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.18.10202-10205.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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