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Journal of Virology, October 2001, p. 9808-9818, Vol. 75, No. 20
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.20.9808-9818.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Cellular COPII Proteins Are Involved in Production of the Vesicles That Form the Poliovirus Replication Complex

René C. Rust,1 Lukas Landmann,2 Rainer Gosert,1 Bor Luen Tang,3 Wanjin Hong,3 Hans-Peter Hauri,4 Denise Egger,1 and Kurt Bienz1,*

Institutes for Medical Microbiology1 and Anatomy2 and Department of Pharmacology, Biocenter,4 University of Basel, CH-4000 Basel, Switzerland, and Institute of Molecular and Cell Biology, University of Singapore, Singapore 1176093

Received 4 April 2001/Accepted 11 July 2001

Poliovirus (PV) replicates its genome in association with membranous vesicles in the cytoplasm of infected cells. To elucidate the origin and mode of formation of PV vesicles, immunofluorescence labeling with antibodies against the viral vesicle marker proteins 2B and 2BC, as well as cellular markers of the endoplasmic reticulum (ER), anterograde transport vesicles, and the Golgi complex, was performed in BT7-H cells. Optical sections obtained by confocal laser scanning microscopy were subjected to a deconvolution process to enhance resolution and signal-to-noise ratio and to allow for a three-dimensional representation of labeled membrane structures. The mode of formation of the PV vesicles was, on morphological grounds, similar to the formation of anterograde membrane traffic vesicles in uninfected cells. ER-resident membrane markers were excluded from both types of vesicles, and the COPII components Sec13 and Sec31 were both found to be colocalized on the vesicular surface, indicating the presence of a functional COPII coat. PV vesicle formation during early time points of infection did not involve the Golgi complex. The expression of PV protein 2BC or the entire P2 and P3 genomic region led to the production of vesicles carrying a COPII coat and showing the same mode of formation as vesicles produced after PV infection. These results indicate that PV vesicles are formed at the ER by the cellular COPII budding mechanism and thus are homologous to the vesicles of the anterograde membrane transport pathway.

* 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 3283. E-mail: Kurt.Bienz{at}unibas.ch.

Journal of Virology, October 2001, p. 9808-9818, Vol. 75, No. 20
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.20.9808-9818.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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