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Entry of dengue virus 2 (DENV-2) into Aedes albopictus mosquito C6/36 cells was analysed using biochemical and molecular inhibitors, together with confocal and electron microscopy observations. Treatment with monodansylcadaverine, chlorpromazine, sucrose and ammonium chloride inhibited DENV-2 virus yield and protein expression, whereas nystatin, a blocker of caveolae-mediated endocytosis, did not have any effect. Using confocal microscopy, co-localization of DENV-2 E glycoprotein and the marker protein transferrin was observed at the periphery of the cytoplasm. To support the requirement of clathrin function for DENV-2 entry, overexpression of a dominant-negative mutant of Eps15 in C6/36 cells was shown to impair virus entry. The disruption of actin microfilaments by cytochalasin D also significantly affected DENV-2 replication. In contrast, microtubule disruption by colchicine treatment did not impair DENV-2 infectivity, suggesting that DENV-2 does not require transport from early to late endosomes for successful infection of mosquito cells. Furthermore, using transmission electron microscopy, DENV-2 particles of approximately 44-52 nm were found attached within electron-dense invaginations of the plasma membrane and in coated vesicles that resembled those of clathrin-coated pits and vesicles, respectively. Together, these results demonstrate for the first time that DENV-2 enters insect cells by receptor-mediated, clathrin-dependent endocytosis, requiring traffic through an acidic pH compartment for subsequent uncoating and completion of a productive infection. © 2008 SGM.


Documento: Artículo
Título:Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis
Autor:Acosta, E.G.; Castilla, V.; Damonte, E.B.
Filiación:Laboratorio de Virología, Departamento de Química Biológica, Universidad de Buenos Aires, 1428 Buenos Aires, Argentina
Palabras clave:actin; ammonium chloride; chlorpromazine; clathrin; colchicine; cytochalasin D; dansylcadaverine; nystatin; sucrose; transferrin; virus glycoprotein; clathrin; cytochalasin D; virus envelope protein; Aedes albopictus; animal cell; article; cell membrane; coated pit; coated vesicle; confocal microscopy; controlled study; Dengue virus; dengue virus 2; electron microscopy; endocytosis; endosome; Flavivirus; microtubule; mosquito; nonhuman; pH; priority journal; protein expression; protein localization; virus entry; virus infectivity; virus particle; virus replication; Aedes; animal; Cercopithecus; drug effect; genetics; metabolism; microfilament; physiology; transport at the cellular level; ultrastructure; Vero cell; virology; Aedes albopictus; Dengue virus; Hexapoda; Actins; Aedes; Animals; Biological Transport; Cercopithecus aethiops; Clathrin; Cytochalasin D; Dengue Virus; Endocytosis; Endosomes; Microfilaments; Microscopy, Confocal; Microtubules; Transferrin; Vero Cells; Viral Envelope Proteins; Virus Internalization
Página de inicio:474
Página de fin:484
Título revista:Journal of General Virology
Título revista abreviado:J. Gen. Virol.
CAS:ammonium chloride, 12125-02-9; chlorpromazine, 50-53-3, 69-09-0; colchicine, 64-86-8; cytochalasin D, 22144-77-0; dansylcadaverine, 10121-91-2; nystatin, 1400-61-9, 34786-70-4, 62997-67-5; sucrose, 122880-25-5, 57-50-1; transferrin, 82030-93-1; Actins; Clathrin; Cytochalasin D, 22144-77-0; Transferrin, 11096-37-0; Viral Envelope Proteins


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---------- APA ----------
Acosta, E.G., Castilla, V. & Damonte, E.B. (2008) . Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis. Journal of General Virology, 89(2), 474-484.
---------- CHICAGO ----------
Acosta, E.G., Castilla, V., Damonte, E.B. "Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis" . Journal of General Virology 89, no. 2 (2008) : 474-484.
---------- MLA ----------
Acosta, E.G., Castilla, V., Damonte, E.B. "Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis" . Journal of General Virology, vol. 89, no. 2, 2008, pp. 474-484.
---------- VANCOUVER ----------
Acosta, E.G., Castilla, V., Damonte, E.B. Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis. J. Gen. Virol. 2008;89(2):474-484.