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Journal of Virology, January 1999, p. 427-435, Vol. 73, No. 1
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Functional Coupling between Replication and Packaging of Poliovirus Replicon RNA

Constance I. Nugent,1,dagger Kyle L. Johnson,2,Dagger Peter Sarnow,2,3 and Karla Kirkegaard1,3,*

Department of Molecular, Cellular and Developmental Biology and Howard Hughes Medical Institute, University of Colorado, Boulder, Colorado 803091; Department of Biochemistry, Biophysics and Genetics, University of Colorado Health Sciences Center, Denver, Colorado 802622; and Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California 943053

Received 22 May 1998/Accepted 16 September 1998

Poliovirus RNA genomes that contained deletions in the capsid-coding regions were synthesized in monkey kidney cells that constitutively expressed T7 RNA polymerase. These replication-competent subgenomic RNAs, or replicons (G. Kaplan and V. R. Racaniello, J. Virol. 62:1687-1696, 1988), were encapsidated in trans by superinfecting polioviruses. When superinfecting poliovirus resistant to the antiviral compound guanidine was used, the RNA replication of the replicon RNAs could be inhibited independently of the RNA replication of the guanidine-resistant helper virus. Inhibiting the replication of the replicon RNA also profoundly inhibited its trans-encapsidation, even though the capsid proteins present in the cells could efficiently encapsidate the helper virus. The observed coupling between RNA replication and RNA packaging could account for the specificity of poliovirus RNA packaging in infected cells and the observed effects of mutations in the coding regions of nonstructural proteins on virion morphogenesis. It is suggested that this coupling results from direct interactions between the RNA replication machinery and the capsid proteins. The coupling of RNA packaging to RNA replication and of RNA replication to translation (J. E. Novak and K. Kirkegaard, Genes Dev. 8:1726-1737, 1994) could serve as mechanisms for late proofreading of poliovirus RNA, allowing only those RNA genomes capable of translating a full complement of functional RNA replication proteins to be propagated.


* Corresponding author. Mailing address: Department of Microbiology and Immunology, Stanford University School of Medicine, Sherman Fairchild Science Bldg., 299 Campus Dr., Stanford, CA 94305-5124. Phone: (650) 498-7075. Fax: (650) 498-7147. E-mail: karlak{at}leland.stanford.edu.

dagger Present address: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030.

Dagger Present address: Department of Microbiology, University of Alabama, Birmingham, AL 35294.


Journal of Virology, January 1999, p. 427-435, Vol. 73, No. 1
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.



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