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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,
Kyle L.
Johnson,2,
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.

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

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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