info:eu-repo/semantics/article
Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis
Fecha
2008-02Registro en:
Acosta, Eliana Gisela; Castilla, Viviana; Damonte, Elsa Beatriz; Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis; Society for General Microbiology; Journal of General Virology; 89; 2; 2-2008; 474-484
0022-1317
CONICET Digital
CONICET
Autor
Acosta, Eliana Gisela
Castilla, Viviana
Damonte, Elsa Beatriz
Resumen
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.