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Journal of Virology, September 2008, p. 8838-8848, Vol. 82, No. 17
0022-538X/08/$08.00+0     doi:10.1128/JVI.00398-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Association of Hepatitis C Virus Replication Complexes with Microtubules and Actin Filaments Is Dependent on the Interaction of NS3 and NS5A{triangledown}

Chao-Kuen Lai,1 King-Song Jeng,1 Keigo Machida,2 and Michael M. C. Lai1,2,3*

Institute of Molecular Biology, Academia Sinica, Taipei 115, Taiwan,1 Department of Molecular Microbiology and Immunology, University of Southern California Keck School of Medicine, 2001 Zonal Avenue, Los Angeles, California, 90033,2 National Cheng Kung University, Tainan 701, Taiwan3

Received 25 February 2008/ Accepted 6 June 2008

The hepatitis C virus (HCV) RNA replication complex (RC), which is composed of viral nonstructural (NS) proteins and host cellular proteins, replicates the viral RNA genome in association with intracellular membranes. Two viral NS proteins, NS3 and NS5A, are essential elements of the RC. Here, by using immunoprecipitation and fluorescence resonance energy transfer assays, we demonstrated that NS3 and NS5A interact with tubulin and actin. Furthermore, immunofluorescence microscopy and electron microscopy revealed that HCV RCs were aligned along microtubules and actin filaments in both HCV replicon cells and HCV-infected cells. In addition, the movement of RCs was inhibited when microtubules or actin filaments were depolymerized by colchicine and cytochalasin B, respectively. Based on our observations, we propose that microtubules and actin filaments provide the tracks for the movement of HCV RCs to other regions in the cell, and the molecular interactions between RCs and microtubules, or RCs and actin filaments, are mediated by NS3 and NS5A.

* Corresponding author. Mailing address: Institute of Molecular Biology, Academia Sinica, Taipei 115, Taiwan. Phone: 886-2-27892365. Fax: 886-2-27826085. E-mail: michlai{at}gate.sinica.edu.tw

{triangledown} Published ahead of print on 18 June 2008.

Journal of Virology, September 2008, p. 8838-8848, Vol. 82, No. 17
0022-538X/08/$08.00+0     doi:10.1128/JVI.00398-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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