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標題: Bovine Ephemeral Fever Virus Uses a Clathrin-Mediated and Dynamin 2-Dependent Endocytosis Pathway That Requires Rab5 and Rab7 as Well as Microtubules
作者: Ching, Y.Cheng
Wing, L.Shih
Wei, R.Huang
Pei, I.Chi
Ming, H.Wu
Hung, J.Liu
Project: Journal of Virology, Volume 86, Issue 24, page(s) 13653–13661.
The specific cell pathways involved in bovine ephemeral fever virus (BEFV) cell entry have not been determined. In this work,
colocalization of theMprotein of BEFV with clathrin or dynamin 2 was observed under a fluorescence microscope. To better
understand BEFV entry, we carried out internalization studies with a fluorescently labeled BEFV by using a lipophilic dye, 3,30-
dilinoleyloxacarbocyanine perchlorate (DiO), further suggesting that BEFV uses a clathrin-mediated endocytosis pathway. Our
results suggest that clathrin-mediated and dynamin 2-dependent endocytosis is an important avenue of BEFV entry. Suppression
of Rab5 or Rab7a through the use of a Rab5 dominant negative mutant and Rab7a short hairpin RNA (shRNA) demonstrated
that BEFV requires both early and late endosomes for endocytosis and subsequent infection in MDBK and Vero cells.
Treatment of BEFV-infected cells with nocodazole significantly decreased theMprotein synthesis and viral yield, indicating that
microtubules play an important role in BEFV productive infection, likely by mediating trafficking of BEFV-containing endosomes.
Furthermore, BEFV infection was strongly blocked by different inhibitors of endosomal acidification, suggesting that
virus enters host cells by clathrin-mediated and dynamin 2-dependent endocytosis in a pH-dependent manner.
DOI: 10.1128/JVI.01073-12
Appears in Collections:分子生物學研究所

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