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Journal of Virology, February 2007, p. 1140-1147, Vol. 81, No. 3
0022-538X/07/$08.00+0     doi:10.1128/JVI.01345-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Vaccinia Virus Interactions with the Cell Membrane Studied by New Chromatic Vesicle and Cell Sensor Assays{triangledown}

Z. Orynbayeva,1 S. Kolusheva,1 N. Groysman,1 N. Gavrielov,2 L. Lobel,2 and R. Jelinek1*

Department of Chemistry and Ilse Katz Center for Nanotechnology,1 Department of Virology, Ben Gurion University of the Negev, Beer Sheva, Israel 841052

Received 26 June 2006/ Accepted 6 November 2006

The potential danger of cross-species viral infection points to the significance of understanding the contributions of nonspecific membrane interactions with the viral envelope compared to receptor-mediated uptake as a factor in virus internalization and infection. We present a detailed investigation of the interactions of vaccinia virus particles with lipid bilayers and with epithelial cell membranes using newly developed chromatic biomimetic membrane assays. This analytical platform comprises vesicular particles containing lipids interspersed within reporter polymer units that emit intense fluorescence following viral interactions with the lipid domains. The chromatic vesicles were employed as membrane models in cell-free solutions and were also incorporated into the membranes of epithelial cells, thereby functioning as localized membrane sensors on the cell surface. These experiments provide important insight into membrane interactions with and fusion of virions and the kinetic profiles of these processes. In particular, the data emphasize the significance of cholesterol/sphingomyelin domains (lipid rafts) as a crucial factor promoting bilayer insertion of the viral particles. Our analysis of virus interactions with polymer-labeled living cells exposed the significant role of the epidermal growth factor receptor in vaccinia virus infectivity; however, the data also demonstrated the existence of additional non-receptor-mediated mechanisms contributing to attachment of the virus to the cell surface and its internalization.

* Corresponding author. Mailing address: Department of Chemistry, Ben Gurion University, Beer Sheva, Israel 84103. Phone: 972-8-6461747. Fax: 972-8-6472943. E-mail: razj{at}bgu.ac.il.

{triangledown} Published ahead of print on 15 November 2006.

Journal of Virology, February 2007, p. 1140-1147, Vol. 81, No. 3
0022-538X/07/$08.00+0     doi:10.1128/JVI.01345-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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