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Journal of Virology, July 2000, p. 6570-6580, Vol. 74, No. 14
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Formation of the Poliovirus Replication Complex Requires Coupled
Viral Translation, Vesicle Production, and Viral RNA
Synthesis
Denise
Egger,1
Natalya
Teterina,2
Ellie
Ehrenfeld,2 and
Kurt
Bienz1,*
Institute for Medical Microbiology,
University of Basel, Basel, Switzerland,1 and
Laboratory of Viral Diseases, National Institute of Allergy
and Infectious Diseases, National Institutes of Health, Bethesda,
Maryland2
Received 23 December 1999/Accepted 17 April 2000
Poliovirus (PV) infection induces the rearrangement of
intracellular membranes into characteristic vesicles which assemble into an RNA replication complex. To investigate this transformation, endoplasmic reticulum (ER) membranes in HeLa cells were modified by the
expression of different cellular or viral membrane-binding proteins.
The membrane-binding proteins induced two types of membrane alterations, i.e., extended membrane sheets and vesicles similar to
those found during a PV infection. Cells expressing membrane-binding proteins were superinfected with PV and then analyzed for virus replication, location of membranes, viral protein, and RNA by immunofluorescence and fluorescent in situ hybridization. Cultures expressing cellular or viral membrane-binding proteins, but not those
expressing soluble proteins, showed a markedly reduced ability to
support PV replication as a consequence of the modification of ER
membranes. The altered membranes, regardless of their morphology, were
not used for the formation of viral replication complexes during a
subsequent PV infection. Specifically, membrane sheets were not
substrates for PV-induced vesicle formation, and, surprisingly, vesicles induced by and carrying one or all of the PV replication proteins did not contribute to replication complexes formed by the
superinfecting PV. The formation of replication complexes required
active viral RNA replication. The extensive alterations induced by
membrane-binding proteins in the ER resulted in reduced viral protein
synthesis, thus affecting the number of cells supporting PV
multiplication. Our data suggest that a functional replication complex
is formed in cis, in a coupled process involving viral translation, membrane modification and vesicle budding, and viral RNA synthesis.
*
Corresponding author. Mailing address: Institute for
Medical Microbiology, University of Basel, Petersplatz 10, CH-4003
Basel, Switzerland. Phone: 41 61 267 3290. Fax: 41 61 267 3298. E-mail: Kurt.Bienz{at}unibas.ch.
Journal of Virology, July 2000, p. 6570-6580, Vol. 74, No. 14
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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