Cellular origin and ultrastructure of membranes induced during poliovirus infection
Poliovirus RNA replicative complexes are associated with cytoplasmic membranous structures that accumulate during viral infection. These membranes were immunoisolated by using a monoclonal antibody against the viral nonstructural protein 2C. Biochemical analysis of the isolated membranes revealed that several organelles of the host cell (lysosomes, trans-Golgi stack and trans-Golgi network, and endoplasmic reticulum) contributed to the virus-induced membranous structures. Electron microscopy of infected cells preserved by high-pressure freezing revealed that the virus-induced membranes contain double lipid bilayers that surround apparently cytosolic material. Immunolabeling experiments showed that poliovirus proteins 2C and 3D were localized to the same membranes as the cellular markers tested. The morphological and biochemical data are consistent with the hypothesis that autophagy or a similar host process is involved in the formation of the poliovirus-induced membranes.
© 1996 American Society for Microbiology. Received 2 February 1996; Accepted 8 June 1996. We are indebted to Kurt Bienz and Denise Egger for their generous gift of anti-2C monoclonal antibody. We thank L. Andrew Staehelin and Kathryn Howell for their insights into cytoplasmic membrane morphology and Anne McBride and Frank Luca for their roles in the preparation of the anti-3D monoclonal antibody. We are grateful to Peter Sarnow, Kurt Bienz, John R. Doedens, Denise Egger, Anne E. McBride, and L. Andrew Staehelin for helpful comments on the manuscript and to Steven Fuller, Hans-Peter Hauri, Eric Berger, and Minoru Fukuda for the provision of antibodies critical to this research. A.S. was a postdoctoral research associate, and K.K. was an assistant investigator, of the Howard Hughes Medical Institute. A.S. also acknowledges support of the Swiss National Science Foundation. This work was supported by the Howard Hughes Medical Institute and by NIH grant AI-25166.
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