Initiation of duck hepatitis B virus infection requires cleavage by a furin-like protease.
The entry mechanism of hepatitis B virus (HBV) has not been defined, and this impedes development of antiviral therapies aimed at an early step in the viral life cycle. HBV infection has both host and tissue specificities. For the related duck hepatitis B virus (DHBV), duck carboxypeptidase D (DCPD) has been proposed as the species-specific docking receptor, while glycine decarboxylase (DGD) may serve as a tissue-specific cofactor or secondary receptor. DGD binds to several truncated versions of the viral large envelope protein but not to the full-length protein, suggesting a need for proteolytic cleavage of the envelope protein by a furin-like proprotein convertase. In the present study, we found that transfected DCPD could confer DHBV binding to non-duck cell lines but that this was followed by rapid virus release from cells. Coexpression of furin led to DCPD cleavage and increased virus retention. Treatment of DHBV particles with endosome prepared from duck liver led to cleavage of the large envelope protein, and such viral preparation could generate a small amount of covalently closed circular DNA in LMH cells, a chicken hepatoma cell line resistant to DHBV infection. A furin inhibitor composed of decanoyl-RVKR-chloromethylketone blocked endosomal cleavage of the large envelope protein in vitro and suppressed DHBV infection of primary duck hepatocytes in vivo. These findings suggest that furin or a furin-like proprotein convertase facilitates DHBV infection by cleaving both the docking receptor and the viral large envelope protein.
Tong
Yupin
Y
Liver Research Center, Rhode Island Hospital and Warren Alpert Medical School of Brown University, Providence, Rhode Island 02903, USA.
Tong
Shuping
S
Zhao
Xiaoai
X
Wang
Jianguo
J
Jun
Jenny
J
Park
Joseph
J
Wands
Jack
J
Li
Jisu
J
D000818:Animals; D002460:Cell Line; D002645:Chickens; D004372:Ducks; D045683:Furin / Q000378:metabolism*; D015611:Hepatitis B Virus, Duck / Q000502:physiology*; D022781:Hepatocytes / Q000821:virology; D014759:Viral Envelope Proteins / Q000378:metabolism*; D053586:Virus Internalization
D014759:Viral Envelope Proteins; D045683:Furin