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Journal of Virology, March 2001, p. 2119-2129, Vol. 75, No. 5
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.5.2119-2129.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Self-Assembly of Nucleocapsid-Like Particles from Recombinant Hepatitis C Virus Core Protein

Meghan Kunkel,1 Marta Lorinczi,1 René Rijnbrand,2 Stanley M. Lemon,2 and Stanley J. Watowich1,*

Department of Human Biological Chemistry & Genetics and the Sealy Center for Structural Biology1 and Department of Microbiology and Immunology,2 University of Texas Medical Branch, Galveston, Texas 77555

Received 10 August 2000/Accepted 30 November 2000

Little is known about the assembly pathway and structure of hepatitis C virus (HCV) since insufficient quantities of purified virus are available for detailed biophysical and structural studies. Here, we show that bacterially expressed HCV core proteins can efficiently self-assemble in vitro into nucleocapsid-like particles. These particles have a regular, spherical morphology with a modal distribution of diameters of approximately 60 nm. Self-assembly of nucleocapsid-like particles requires structured RNA molecules. The 124 N-terminal residues of the core protein are sufficient for self-assembly into nucleocapsid-like particles. Inclusion of the carboxy-terminal domain of the core protein modifies the core assembly pathway such that the resultant particles have an irregular outline. However, these particles are similar in size and shape to those assembled from the 124 N-terminal residues of the core protein. These results provide novel opportunities to delineate protein-protein and protein-RNA interactions critical for HCV assembly, to study the molecular details of HCV assembly, and for performing high-throughput screening of assembly inhibitors.

* Corresponding author. Mailing address: Department of Human Biological Chemistry & Genetics, University of Texas Medical Branch, Galveston, TX 77555-0645. Phone: (409) 747-4749. Fax: (409) 747-4745. E-mail: watowich{at}bloch.utmb.edu.

Journal of Virology, March 2001, p. 2119-2129, Vol. 75, No. 5
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.5.2119-2129.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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