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Journal of Virology, October 1998, p. 7972-7977, Vol. 72, No. 10
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

trans-Encapsidation of a Poliovirus Replicon by Different Picornavirus Capsid Proteins

Xi-Yu Jia,1 Marc Van Eden,1 Marc G. Busch,1 Ellie Ehrenfeld,2 and Donald F. Summers1,*

Departments of Microbiology and Molecular Genetics1 and of Molecular Biology and Biochemistry,2 University of California, Irvine, California 92697

Received 15 December 1997/Accepted 1 July 1998

A trans-encapsidation assay was established to study the specificity of picornavirus RNA encapsidation. A poliovirus replicon with the luciferase gene replacing the capsid protein-coding region was coexpressed in transfected HeLa cells with capsid proteins from homologous or heterologous virus. Successful trans-encapsidation resulted in assembly and production of virions whose replication, upon subsequent infection of HeLa cells, was accompanied by expression of luciferase activity. The amount of luciferase activity was proportional to the amount of trans-encapsidated virus produced from the cotransfection. When poliovirus capsid proteins were supplied in trans, >2 × 106 infectious particles/ml were produced. When coxsackievirus B3, human rhinovirus 14, mengovirus, or hepatitis A virus (HAV) capsid proteins were supplied in trans, all but HAV showed some encapsidation of the replicon. The overall encapsidation efficiency of the replicon RNA by heterologous capsid proteins was significantly lower than when poliovirus capsid was used. trans-encapsidated particles could be completely neutralized with specific antisera against each of the donor virus capsids. The results indicate that encapsidation is regulated by specific viral nucleic acid and protein sequences.

* Corresponding author. Present address: NCI, NIH, Frederick Cancer Research Center, P.O. Box B, Bldg. 427, Room 10, Frederick, MD 21702-1124. Phone: (301) 846-5096. Fax: (301) 846-1494. E-mail: dfsummer{at}ncifcrf.gov.

Journal of Virology, October 1998, p. 7972-7977, Vol. 72, No. 10
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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