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, pHdependent 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 dynaminindependent in Hep3B/miR122 cells. Collectively, these data indicated that HCV entered different target cells through different entry routes.
| Original language | English |
|---|---|
| Pages (from-to) | 2658-2667 |
| Number of pages | 10 |
| Journal | Journal of General Virology |
| Volume | 95 |
| DOIs | |
| Publication status | Published - Dec 1 2014 |
Fingerprint
ASJC Scopus subject areas
- Virology
- Medicine(all)
Cite this
Alternative endocytosis pathway for productive entry of hepatitis C virus. / Matsuda, Mami; Suzuki, Ryosuke; Kataoka, Chikako; Watashi, Koichi; Aizaki, Hideki; Kato, Nobuyuki; Matsuura, Yoshiharu; Suzuki, Tetsuro; Wakita, Takaji.
In: Journal of General Virology, Vol. 95, 01.12.2014, p. 2658-2667.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Alternative endocytosis pathway for productive entry of hepatitis C virus
AU - Matsuda, Mami
AU - Suzuki, Ryosuke
AU - Kataoka, Chikako
AU - Watashi, Koichi
AU - Aizaki, Hideki
AU - Kato, Nobuyuki
AU - Matsuura, Yoshiharu
AU - Suzuki, Tetsuro
AU - Wakita, Takaji
PY - 2014/12/1
Y1 - 2014/12/1
N2 - 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, pHdependent 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 dynaminindependent in Hep3B/miR122 cells. Collectively, these data indicated that HCV entered different target cells through different entry routes.
AB - 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, pHdependent 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 dynaminindependent in Hep3B/miR122 cells. Collectively, these data indicated that HCV entered different target cells through different entry routes.
UR - http://www.scopus.com/inward/record.url?scp=84910597301&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84910597301&partnerID=8YFLogxK
U2 - 10.1099/vir.0.068528-0
DO - 10.1099/vir.0.068528-0
M3 - Article
C2 - 25096815
AN - SCOPUS:84910597301
VL - 95
SP - 2658
EP - 2667
JO - Journal of General Virology
JF - Journal of General Virology
SN - 0022-1317
ER -