Interaction of West Nile virus with alpha v beta 3 integrin mediates virus entry into cells.
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.
Flavivirology Laboratory, Department of Microbiology, 5 Science Drive 2, National University of Singapore, Singapore 117597.
Justin Jang-Hann
JJ
Chu
Mah-Lee
ML
Ng
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001665:Binding Sites; D002413:Cations, Divalent / Q000494:pharmacology; D002460:Cell Line; D002642:Chick Embryo; D002522:Chlorocebus aethiops; D006224:Cricetinae; D015870:Gene Expression; D020868:Gene Silencing; D006367:HeLa Cells; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D039661:Integrin beta3 / Q000235:genetics / Q000502:physiology; D007668:Kidney; D008545:Melanoma; D009842:Oligopeptides / Q000378:metabolism / Q000494:pharmacology; D011991:Receptors, Virus / Q000235:genetics / Q000502:physiology*; D011994:Recombinant Proteins; D015398:Signal Transduction; D012995:Solubility; D014162:Transfection; D014407:Tumor Cells, Cultured; D014709:Vero Cells; D014902:West Nile virus / Q000502:physiology*
D000906:Antibodies; D002413:Cations, Divalent; D039302:Integrin alphaVbeta3; D039661:Integrin beta3; D009842:Oligopeptides; D011991:Receptors, Virus; D011994:Recombinant Proteins; C047981:arginyl-glycyl-aspartic acid