Abstract
Influenza A virus H5N1 presents a major threat to human health. The entry of influenza virus into host cells is believed to be mediated by hemagglutinin (HA), a virus surface glycoprotein that can bind terminal sialic acid residues on host cell glycoproteins and glycolipids. In this study, we elucidated the pathways through which H5N1 enters human lung carcinoma cell line A549. We first proved that H5N1 can enter A549 cells via endocytosis, as lysosomotropic agents, such as bafilomycin A1 and chloroquine, can rescue H5N1-induced A549 cell death. By using specific inhibitors, and siRNAs that target the clathrin pathway, we further found that H5N1 could enter A549 cells via clathrin-mediated endocytosis, while inhibitors targeting caveolae-mediated endocytosis could not inhibit H5N1 cell entry. These findings expand our understanding of H5N1 pathogenesis and provide new information for anti-viral drug research.
Article PDF
Similar content being viewed by others
References
Isolation of avian influenza A(H5N1) viruses from humans—Hong Kong, May–December 1997. MMWR Morb Mortal Wkly Rep, 1997, 46: 1204–1207
Allen P J. Avian influenza pandemic: not if, but when. Pediatr Nurs, 2006, 32: 76–81, 16572544
Pascoe N. A pandemic flu: not if, but when. SARS was the wake-up call we slept through. Tex Nurs, 2006, 80: 6–10, 16681227
Sieczkarski S B, Whittaker G R. Dissecting virus entry via endocytosis. J Gen Virol, 2002, 83: 1535–1545, 12075072, 1:CAS:528:DC%2BD38Xlt1Wgurk%3D
Nabi I R, Le P U. Caveolae/raft-dependent endocytosis. J Cell Biol, 2003, 161: 673–677, 12771123, 10.1083/jcb.200302028, 1:CAS:528:DC%2BD3sXktlWiur4%3D
Marsh M, Helenius A. Virus entry: open sesame. Cell, 2006, 124: 729–740, 16497584, 10.1016/j.cell.2006.02.007, 1:CAS:528:DC%2BD28Xit1KltLk%3D
Pelkmans L, Helenius A. Insider information: what viruses tell us about endocytosis. Curr Opin Cell Biol, 2003, 15: 414–422, 12892781, 10.1016/S0955-0674(03)00081-4, 1:CAS:528:DC%2BD3sXlvVGjsbc%3D
Matlin K S, Reggio H, Helenius A, et al. Infectious entry pathway of influenza virus in a canine kidney cell line. J Cell Biol, 1981, 91: 601–613, 7328111, 10.1083/jcb.91.3.601, 1:STN:280:DyaL387itVamsA%3D%3D
Rust M J, Lakadamyali M, Zhang F, et al. Assembly of endocytic machinery around individual influenza viruses during viral entry. Nat Struct Mol Biol, 2004, 11: 567–573, 15122347, 10.1038/nsmb769, 1:CAS:528:DC%2BD2cXkt1Ogtbs%3D
Nunes-Correia I, Eulalio A, Nir S, et al. Caveolae as an additional route for influenza virus endocytosis in MDCK cells. Cell Mol Biol Lett, 2004, 9: 47–60, 15048150
Sieczkarski S B, Whittaker G R. Influenza virus can enter and infect cells in the absence of clathrin-mediated endocytosis. J Virol, 2002, 76: 10455–10464, 12239322, 10.1128/JVI.76.20.10455-10464.2002, 1:CAS:528:DC%2BD38XnsV2kurk%3D
Wang H, Yang P, Liu K, et al. SARS coronavirus entry into host cells through a novel clathrin- and caveolae-independent endocytic pathway. Cell Res, 2008, 18: 290–301, 18227861, 10.1038/cr.2008.15, 1:CAS:528:DC%2BD1cXhs1OmtLc%3D
Turpin E, Luke K, Jones J, et al. Influenza virus infection increases p53 activity: role of p53 in cell death and viral replication. J Virol, 2005, 79: 8802–8811, 15994774, 10.1128/JVI.79.14.8802-8811.2005, 1:CAS:528:DC%2BD2MXmtFOjs78%3D
Lyles D S. Cytopathogenesis and inhibition of host gene expression by RNA viruses. Microbiol Mol Biol Rev, 2000, 64: 709–724, 11104816, 10.1128/MMBR.64.4.709-724.2000, 1:CAS:528:DC%2BD3MXltFWjtro%3D
Le Roy C, Wrana J L. Clathrin- and non-clathrin-mediated endocytic regulation of cell signalling. Nat Rev Mol Cell Biol, 2005, 6: 112–126, 15687999, 10.1038/nrm1571
Kirchhausen T. Clathrin. Annu Rev Biochem, 2000, 69: 699–727, 10966473, 10.1146/annurev.biochem.69.1.699, 1:CAS:528:DC%2BD3cXnt1ajur0%3D
Wang L H, Rothberg K G, Anderson R G. Mis-assembly of clathrin lattices on endosomes reveals a regulatory switch for coated pit formation. J Cell Biol, 1993, 123: 1107–1117, 8245121, 10.1083/jcb.123.5.1107, 1:CAS:528:DyaK3sXmsVKkt70%3D
Royle S J. The cellular functions of clathrin. Cell Mol Life Sci, 2006, 63: 1823–1832, 16699812, 10.1007/s00018-005-5587-0, 1:CAS:528:DC%2BD28XpsVOls7o%3D
Meertens L, Bertaux C, Dragic T. Hepatitis C virus entry requires a critical postinternalization step and delivery to early endosomes via clathrin-coated vesicles. J Virol, 2006, 80: 11571–11578, 17005647, 10.1128/JVI.01717-06, 1:CAS:528:DC%2BD28Xht12hu7jO
Sun X, Yau V K, Briggs B J, et al. Role of clathrin-mediated endocytosis during vesicular stomatitis virus entry into host cells. Virology, 2005, 338: 53–60, 15936793, 10.1016/j.virol.2005.05.006, 1:CAS:528:DC%2BD2MXlsVChsLs%3D
Stan R V. Structure of caveolae. Biochim Biophys Acta, 2005, 1746: 334–348, 16214243, 10.1016/j.bbamcr.2005.08.008, 1:CAS:528:DC%2BD2MXhtlCqtLjM
Simons K, Toomre D. Lipid rafts and signal transduction. Nat Rev Mol Cell Biol, 2000, 1: 31–39, 11413487, 10.1038/35036052, 1:CAS:528:DC%2BD3MXivVGjtbo%3D
Orlandi P A, Fishman P H. Filipin-dependent inhibition of cholera toxin: evidence for toxin internalization and activation through caveolae-like domains. J Cell Biol, 1998, 141: 905–915, 9585410, 10.1083/jcb.141.4.905, 1:CAS:528:DyaK1cXjt1Kgtbk%3D
Sanchez-San Martin C, Lopez T, Arias C F, et al. Characterization of rotavirus cell entry. J Virol, 2004, 78: 2310–2318, 14963127, 10.1128/JVI.78.5.2310-2318.2004, 1:CAS:528:DC%2BD2cXhs1SltrY%3D
Rejman J, Oberle V, Zuhorn I S, et al. Size-dependent internalization of particles via the pathways of clathrin- and caveolae-mediated endocytosis. Biochem J, 2004, 377: 159–169, 14505488, 10.1042/BJ20031253, 1:CAS:528:DC%2BD2cXis1Wnsg%3D%3D
Ahn A, Gibbons D L, Kielian M. The fusion peptide of Semliki Forest virus associates with sterol-rich membrane domains. J Virol, 2002, 76: 3267–3275, 11884551, 10.1128/JVI.76.7.3267-3275.2002, 1:CAS:528:DC%2BD38Xit1Gqu70%3D
Guo Y, Rumschlag-Booms E, Wang J, et al. Analysis of hemagglutinin-mediated entry tropism of H5N1 avian influenza. Virol J, 2009, 6: 39, 19341465, 10.1186/1743-422X-6-39
Author information
Authors and Affiliations
Corresponding author
Additional information
Supported by the National Natural Science Foundation of China (Grant No. 30623009) and National Basic Research Program of China (Grant No. 2005CB523000)
Rights and permissions
About this article
Cite this article
Wang, H., Jiang, C. Influenza A virus H5N1 entry into host cells is through clathrin-dependent endocytosis. SCI CHINA SER C 52, 464–469 (2009). https://doi.org/10.1007/s11427-009-0061-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11427-009-0061-0