Alternative endocytosis pathway for productive entry of hepatitis C virus.

Abstract

Previous studies have shown that hepatitis C virus (HCV) enters human hepatic cells through interaction with a series of cellular receptors, followed by clathrin-mediated, pH-dependent endocytosis. Here, we investigated the mechanisms of HCV entry into multiple HCV-permissive human hepatocyte-derived cells using trans-complemented HCV particles (HCVtcp). Knockdown of CD81 and claudin-1, or treatment with bafilomycin A1, reduced infection in Huh-7 and Huh7.5.1 cells, suggesting that HCV entered both cell types via receptor-mediated, pH-dependent endocytosis. Interestingly, knockdown of the clathrin heavy chain or dynamin-2 (Dyn2), as well as expression of the dominant-negative form of Dyn2, reduced infection of Huh-7 cells with HCVtcp, whereas infectious entry of HCVtcp into Huh7.5.1 cells was not impaired. Infection of Huh7.5.1 cells with culture-derived HCV (HCVcc) via a clathrin-independent pathway was also observed. Knockdown of caveolin-1, ADP-ribosylation factor 6 (Arf6), flotillin, p21-activated kinase 1 (PAK1) and the PAK1 effector C-terminal binding protein 1 of E1A had no inhibitory effects on HCVtcp infection into Huh7.5.1 cells, thus suggesting that the infectious entry pathway of HCV into Huh7.5.1 cells was not caveolae-mediated, or Arf6- and flotillin-mediated endocytosis and macropinocytosis, but rather may have occurred via an undefined endocytic pathway. Further analysis revealed that HCV entry was clathrin- and dynamin-dependent in ORL8c and HepCD81/miR122 cells, but productive entry of HCV was clathrin- and dynamin-independent in Hep3B/miR122 cells. Collectively, these data indicated that HCV entered different target cells through different entry routes.

• Authors: Mami Matsuda, Ryosuke Suzuki, Chikako Kataoka, Koichi Watashi, Hideki Aizaki, Nobuyuki Kato, Yoshiharu Matsuura, Tetsuro Suzuki, Takaji Wakita
• Journal: The Journal of general virology (J Gen Virol) Vol. 95 Issue Pt 12 Pg. 2658-2667 (Dec 2014) ISSN: 1465-2099 [Electronic] England
• PMID: 25096815 (Publication Type: Journal Article)
• Copyright: © 2014 The Authors.

Topics

• ADP-Ribosylation Factor 6
• Cell Line
• Endocytosis (physiology)
• Gene Expression Regulation (physiology)
• Hepacivirus (physiology)
• Hepatocytes (metabolism, physiology, virology)
• Humans
• Virus Internalization