Journal
VIROLOGY
Volume 513, Issue -, Pages 195-207Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.virol.2017.09.019
Keywords
DsRNA virus; Reovirus; Aquareovirus; Cell entry; Quantum dot; Caveolae/raft-mediated endocytosis
Categories
Funding
- National Natural Science Foundation of China [31372565, 31400139, 31370190, 31672693]
- State Key Laboratory of Freshwater Ecology and Biotechnology [2013FB05]
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Grass carp reovirus (GCRV), a member of the Aquareovirus genus in the Reoviridae family, is considered the most pathogenic aquareovirus. However, its productive viral entry pathways remain largely unclear. Using a combination of quantum dot (QD)-based live-virus tracking and biochemical assays, we found that extraction of cellular membrane cholesterol with methyl-beta-cyclodextrin (M beta CD) and nystatin strongly inhibited the internalization of GCRVs, and supplementation with cholesterol restored viral infection. In addition, the entry of the virus was restrained by genistein, an inhibitor known to block caveolar endocytosis. Subsequent real-time tracking experiments revealed that the QD-labeled GCRV particles were colocalized with caveolin-1, and transfection of cells with dominant-negative mutant (caveolin-1 Y14F) significantly reduced GCRV infection. In contrast, no effects on virus infection were detected when the clathrin-mediated endocytosis or the macro-pinocytosis inhibitors were used. Our results collectively suggest that aquareoviruses can use caveolae/raft-mediated endocytosis as the primary entry pathway to initiate productive infection.
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