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Journal of Virology, April 2005, p. 4557-4567, Vol. 79, No. 8
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.8.4557-4567.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Heat Shock Protein 90 and Heat Shock Protein 70 Are Components of Dengue Virus Receptor Complex in Human Cells

Jorge Reyes-del Valle, Salvador Chávez-Salinas, Fernando Medina, and Rosa M. del Angel*

Departamento de Patología Experimental, Centro de Investigación y de Estudios Avanzados del IPN, Mexico City, Mexico

Received 8 July 2004/ Accepted 3 December 2004

Dengue virus requires the presence of an unidentified cellular receptor on the surface of the host cell. By using a recently published affinity chromatography approach, an 84-kDa molecule, identified as heat shock protein 90 (HSP90) by matrix-assisted laser desorption ionization-time of flight mass spectrometry, was isolated from neuroblastoma and U937 cells. Based on the ability of HSP90 (84 kDa) to interact with HSP70 (74 kDa) on the surface of monocytes during lipopolysaccharide (LPS) signaling and evidence that LPS inhibits dengue virus infection, the presence of HSP70 was demonstrated in affinity chromatography eluates and by pull-down experiments. Infection inhibition assays support the conclusion that HSP90 and HSP70 participate in dengue virus entry as a receptor complex in human cell lines as well as in monocytes/macrophages. Additionally, our results indicate that both HSPs are associated with membrane microdomains (lipid rafts) in response to dengue virus infection. Moreover, methyl-ß-cyclodextrin, a raft-disrupting drug, inhibits dengue virus infection, supporting the idea that cholesterol-rich membrane fractions are important in dengue virus entry.

* Corresponding author. Mailing address: Departamento de Patología Experimental, Centro de Investigación y de Estudios Avanzados del IPN, Av. I.P.N. 2508, Col. San Pedro Zacatenco, Mexico City D.F., C.P. 07360, Mexico. Phone: (5255) 5061-3800, ext. 5647 or 3345. Fax: (5255) 5747-7107. E-mail: rmangel{at}mail.cinvestav.mx.

Journal of Virology, April 2005, p. 4557-4567, Vol. 79, No. 8
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.8.4557-4567.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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