A Real-Time Method to Study DNA Unwinding by Hepatitis C (HCV) NS3 Helicase

Dennis Matlock, PhD

Harding University

Abstract

Hepatitis C Virus (HCV) infects over 170 million persons worldwide (1-3). It is the leading cause of liver disease in the U.S. and is responsible for most liver transplants. Current treatments for this infectious disease are inadequate; therefore new therapies must be developed (4). It is necessary to understand the mechanism of viral replication in order to discover new targets for disruption of viral replication. The focus of this proposal is non-structural protein 3 (NS3) that is encoded by HCV. The NS3 protein exhibits ATPase, helicase, and protease activities.

Using established and published methods by the principal investigator (Dr. Matlock), the interaction of the helicase domain of NS3 (NS3h) with a nucleic acid substrate will be studied. The specific aim of this proposal is to elucidate the mechanism of NS3 binding, unwinding, and translocation along a nucleic acid substrate. The plan of attack is to employ a sensitive, real-time fluorescence method.

Such studies will provide insight into the mechanism of NS3 that to date have been difficult to examine using conventional methods.