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Originally published In Press as doi:10.1074/jbc.M410208200 on October 7, 2004

J. Biol. Chem., Vol. 279, Issue 52, 54533-54541, December 24, 2004
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Interaction of West Nile Virus with {alpha}v{beta}3 Integrin Mediates Virus Entry into Cells*

Justin Jang-hann Chu and Mah-Lee Ng{ddagger}

From the Flavivirology Laboratory, Department of Microbiology, 5 Science Drive 2, National University of Singapore, Singapore 117597

The functional receptor for the flavivirus West Nile (WNV) infection has been characterized in this study with a combination of biochemical and molecular approaches. A 105-kDa protease-sensitive glycoprotein that binds WNV was isolated from the plasma membrane of cells permissive to WNV infection. The protein was subjected to peptide sequencing, and this glycoprotein was identified as a member of the integrin superfamily. Infection of WNV was shown to be markedly inhibited in Vero cells pretreated with blocking antibodies against {alpha}v{beta}3 integrin and its subunits by receptor competition assay. It was also noted that cells pretreated with antibodies against {alpha}v{beta}3 integrin can effectively inhibit flavivirus Japanese encephalitis but to a lesser extent flavivirus dengue infections. West Nile virus entry is independent of divalent cations and is not highly blocked by arginine-glycine-aspartic acid (RGD) peptides, suggesting that the interaction between the virus and {alpha}v{beta}3 integrin is not highly dependent on the classical RGD binding motif. In addition, gene silencing of the {beta}3 integrin subunit in cells has resulted in cells largely resistant to WNV infection. In contrast, expression of recombinant human {beta}3 integrin substantially increased the permissiveness of CS-1 melanoma cells for WNV infection. Soluble {alpha}v{beta}3 integrin can also effectively block WNV infection in a dose-dependent manner. Furthermore, WNV infection also triggered the outside-in signaling pathway via the activation of integrin-associated focal adhesion kinase. The identification of {alpha}v{beta}3 integrin as a receptor for WNV provides insight into virus-receptor interaction, hence creating opportunities in the development of anti-viral strategies against WNV infection.

Received for publication, September 7, 2004 , and in revised form, October 7, 2004.

* This work is supported by the Biomedical Research Council (Singapore), Project No. 01/1/21/18/003. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

{ddagger} To whom correspondence should be addressed. Tel.: 65-8743283; Fax: 65-7766872; E-mail: micngml{at}nus.edu.sg.


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