Role of the asialoglycoprotein receptor in binding and entry of hepatitis C virus structural proteins in cultured human hepatocytes.
We used a baculovirus-based system to prepare structural proteins of hepatitis C virus (HCV) genotype 1a. Binding of this preparation to cultured human hepatic cells was both dose dependent and saturable. This binding was decreased by calcium depletion and was partially prevented by ligands of the asialoglycoprotein receptor (ASGP-R), thyroglobulin, asialothyroglobulin, and antibody against a peptide in the carbohydrate recognition domain of ASGP-R but not preimmune antibody. Uptake by hepatocytes was observed with both radiolabeled and dye-labeled HCV structural proteins. With hepatocytes expressing the hH1 subunit of the ASGP-R fused to green fluorescent protein, we could show by confocal microscopy that dye stain cointernalized with the fusion protein in an area surrounding the nucleus. Internalization was more efficient with a preparation containing p7 than with one that did not. The two preparations bound to transfected 3T3-L1 cells expressing either both (hH1 and hH2) subunits of the ASGP-R (3T3-22Z cells) or both hH1 and a functionally defective variant of hH2 (3T3-24X cells) but not to parental cells. Additionally, uptake of dye-labeled preparation containing p7 was observed with 3T3-22Z cells but not with 3T3-L1 or 3T3-24X cells or with the preparation lacking p7, suggesting that p7 regulates the internalization properties of HCV structural proteins. Our observations suggest that HCV structural proteins bind to and cointernalize with the ASGP-R in cultured human hepatocytes.
Saunier
Bertrand
B
Edison Biotechnology Institute and College of Osteopathic Medicine, Ohio University, Athens 45701, USA. saunier@ohiou.edu
Triyatni
Miriam
M
Ulianich
Luca
L
Maruvada
Padma
P
Yen
Paul
P
Kohn
Leonard D
LD
D016475:3T3 Cells; D000818:Animals; D037263:Asialoglycoprotein Receptor / Q000502:physiology*; D002118:Calcium / Q000494:pharmacology; D002478:Cells, Cultured; D016174:Hepacivirus / Q000737:chemistry* / Q000502:physiology; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D051379:Mice; D014162:Transfection; D015678:Viral Structural Proteins / Q000378:metabolism*
D037263:Asialoglycoprotein Receptor; D015678:Viral Structural Proteins; D002118:Calcium