Dynamin- and Clathrin-dependent Endocytosis in African Swine Fever Virus Entry

Overview

Journal J Virol

Publisher American Society for Microbiology

Date 2009 Nov 27

PMID 19939916

Citations 86

Authors

Bruno Hernaez

Covadonga Alonso

Affiliations

Soon will be listed here.

Abstract

African swine fever virus (ASFV) is a large DNA virus that enters host cells after receptor-mediated endocytosis and depends on acidic cellular compartments for productive infection. The exact cellular mechanism, however, is largely unknown. In order to dissect ASFV entry, we have analyzed the major endocytic routes using specific inhibitors and dominant negative mutants and analyzed the consequences for ASFV entry into host cells. Our results indicate that ASFV entry into host cells takes place by clathrin-mediated endocytosis which requires dynamin GTPase activity. Also, the clathrin-coated pit component Eps15 was identified as a relevant cellular factor during infection. The presence of cholesterol in cellular membranes, but not lipid rafts or caveolae, was found to be essential for a productive ASFV infection. In contrast, inhibitors of the Na(+)/H(+) ion channels and actin polymerization inhibition did not significantly modify ASFV infection, suggesting that macropinocytosis does not represent the main entry route for ASFV. These results suggest a dynamin-dependent and clathrin-mediated endocytic pathway of ASFV entry for the cell types and viral strains analyzed.

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References

Alcami A, Carrascosa A, Vinuela E . The entry of African swine fever virus into Vero cells. Virology. 1989; 171(1):68-75. DOI: 10.1016/0042-6822(89)90511-4. View

Veiga E, Cossart P . The role of clathrin-dependent endocytosis in bacterial internalization. Trends Cell Biol. 2006; 16(10):499-504. PMC: 7126422. DOI: 10.1016/j.tcb.2006.08.005. View

SEVER S, Damke H, Schmid S . Dynamin:GTP controls the formation of constricted coated pits, the rate limiting step in clathrin-mediated endocytosis. J Cell Biol. 2000; 150(5):1137-48. PMC: 2175254. DOI: 10.1083/jcb.150.5.1137. View

Hernaez B, Escribano J, Alonso C . Visualization of the African swine fever virus infection in living cells by incorporation into the virus particle of green fluorescent protein-p54 membrane protein chimera. Virology. 2006; 350(1):1-14. PMC: 7111804. DOI: 10.1016/j.virol.2006.01.021. View

Brodsky F, Chen C, Knuehl C, Towler M, Wakeham D . Biological basket weaving: formation and function of clathrin-coated vesicles. Annu Rev Cell Dev Biol. 2001; 17:517-68. DOI: 10.1146/annurev.cellbio.17.1.517. View

Valdeira M, Bernardes C, Cruz B, Geraldes A . Entry of African swine fever virus into Vero cells and uncoating. Vet Microbiol. 1998; 60(2-4):131-40. DOI: 10.1016/s0378-1135(98)00152-7. View

Amstutz B, Gastaldelli M, Kalin S, Imelli N, Boucke K, Wandeler E . Subversion of CtBP1-controlled macropinocytosis by human adenovirus serotype 3. EMBO J. 2008; 27(7):956-69. PMC: 2323250. DOI: 10.1038/emboj.2008.38. View

Carrascosa J, Carazo J, Carrascosa A, Garcia N, Santisteban A, Vinuela E . General morphology and capsid fine structure of African swine fever virus particles. Virology. 1984; 132(1):160-72. DOI: 10.1016/0042-6822(84)90100-4. View

McCrossan M, Windsor M, Ponnambalam S, Armstrong J, Wileman T . The trans Golgi network is lost from cells infected with African swine fever virus. J Virol. 2001; 75(23):11755-65. PMC: 114761. DOI: 10.1128/JVI.75.23.11755-11765.2001. View

10.

Rodal S, Skretting G, Garred O, Vilhardt F, van Deurs B, Sandvig K . Extraction of cholesterol with methyl-beta-cyclodextrin perturbs formation of clathrin-coated endocytic vesicles. Mol Biol Cell. 1999; 10(4):961-74. PMC: 25220. DOI: 10.1091/mbc.10.4.961. View

11.

Sun X, Yau V, Briggs B, Whittaker G . Role of clathrin-mediated endocytosis during vesicular stomatitis virus entry into host cells. Virology. 2005; 338(1):53-60. DOI: 10.1016/j.virol.2005.05.006. View

12.

Macia E, Ehrlich M, Massol R, Boucrot E, Brunner C, Kirchhausen T . Dynasore, a cell-permeable inhibitor of dynamin. Dev Cell. 2006; 10(6):839-50. DOI: 10.1016/j.devcel.2006.04.002. View

13.

Raghu H, Sharma-Walia N, Veettil M, Sadagopan S, Chandran B . Kaposi's sarcoma-associated herpesvirus utilizes an actin polymerization-dependent macropinocytic pathway to enter human dermal microvascular endothelial and human umbilical vein endothelial cells. J Virol. 2009; 83(10):4895-911. PMC: 2682094. DOI: 10.1128/JVI.02498-08. View

14.

Sieczkarski S, Whittaker G . Influenza virus can enter and infect cells in the absence of clathrin-mediated endocytosis. J Virol. 2002; 76(20):10455-64. PMC: 136567. DOI: 10.1128/jvi.76.20.10455-10464.2002. View

15.

Helenius A, Kartenbeck J, Simons K, Fries E . On the entry of Semliki forest virus into BHK-21 cells. J Cell Biol. 1980; 84(2):404-20. PMC: 2110562. DOI: 10.1083/jcb.84.2.404. View

16.

Mercer J, Helenius A . Virus entry by macropinocytosis. Nat Cell Biol. 2009; 11(5):510-20. DOI: 10.1038/ncb0509-510. View

17.

Hybiske K, Stephens R . Mechanisms of Chlamydia trachomatis entry into nonphagocytic cells. Infect Immun. 2007; 75(8):3925-34. PMC: 1952008. DOI: 10.1128/IAI.00106-07. View

18.

Kirchhausen T, Macia E, Pelish H . Use of dynasore, the small molecule inhibitor of dynamin, in the regulation of endocytosis. Methods Enzymol. 2008; 438:77-93. PMC: 2796620. DOI: 10.1016/S0076-6879(07)38006-3. View

19.

Damke H, Baba T, van der Bliek A, Schmid S . Clathrin-independent pinocytosis is induced in cells overexpressing a temperature-sensitive mutant of dynamin. J Cell Biol. 1995; 131(1):69-80. PMC: 2120592. DOI: 10.1083/jcb.131.1.69. View

20.

Gomez-Puertas P, Rodriguez F, Oviedo J, Brun A, Alonso C, Escribano J . The African swine fever virus proteins p54 and p30 are involved in two distinct steps of virus attachment and both contribute to the antibody-mediated protective immune response. Virology. 1998; 243(2):461-71. DOI: 10.1006/viro.1998.9068. View