Entry of dengue virus serotype 2 into ECV304 cells depends on clathrin-dependent endocytosis, but not on caveolae-dependent endocytosis.
Caveolae- and clathrin-mediated endocytosis are major internalization pathways used by several pathogens; however, their distinctive roles in dengue virus (DV) entry have not been addressed. In this study, we compared the involvement of caveolae- and clathrin-mediated endocytosis in the infectious entry of DV serotype 2 (DV2) into human endothelial-like ECV304 cells. Confocal microscopy study on DV2-infected cells showed that viral antigens were co-localized with clathrin heavy chains, epidermal growth factor pathway substrate clone 15 (Eps15), and adaptin-alpha, but not with caveolin-1. Treatment with chlorpromazine, which inhibits clathrin-dependent endocytosis, led to reduced virus entry into cells, whereas treatment with nystatin, a caveolae inhibitory agent, did not. Furthermore, gene silencing of Eps15 resulted in an average of 75% reduced infection of ECV304 cells by DV2. Our results demonstrated that DV2 enters ECV304 cells by clathrin-dependent endocytosis, not by caveolae-dependent endocytosis.
Peng
Tao
T
Department of Microbiology, Third Military Medical University, Chongqing, People's Republic of China.
Wang
Jia-Li
JL
Chen
Wei
W
Zhang
Jun-Lei
JL
Gao
Na
N
Chen
Zong-Tao
ZT
Xu
Xiao-Feng
XF
Fan
Dong-Ying
DY
An
Jing
J
D021941:Caveolae / Q000502:physiology*; D002460:Cell Line; D002966:Clathrin / Q000378:metabolism*; D003715:Dengue / Q000378:metabolism / Q000503:physiopathology* / Q000821:virology; D003716:Dengue Virus / Q000502:physiology*; D004705:Endocytosis; D006801:Humans; D053586:Virus Internalization
D002966:Clathrin