Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1995 Nov;69(11):7106–7112. doi: 10.1128/jvi.69.11.7106-7112.1995

Interaction between duck hepatitis B virus and a 170-kilodalton cellular protein is mediated through a neutralizing epitope of the pre-S region and occurs during viral infection.

S Tong 1, J Li 1, J R Wands 1
PMCID: PMC189630  PMID: 7474130

Abstract

Identification of cell surface viral binding proteins is important for understanding viral attachment and internalization. We have fused the pre-S domain of the duck hepatitis B virus (DHBV) large envelope protein to glutathione S-transferase and demonstrated a 170-kDa binding protein (p170) in [35S]methionine-labeled duck hepatocyte lysates. This glycoprotein was found abundantly in all extrahepatic tissues infectible with DHBV and in some noninfectible tissues, though it is not secreted into the blood. The interaction of pre-S fusion protein with p170 was competitively inhibited by wild-type DHBV in a dose-dependent manner. In addition, infection of hepatocytes with DHBV blocked the binding of pre-S fusion protein to p170, which suggests a biological role for p170 during natural infection. The p170 binding site was mapped to a conserved sequence of 16 amino acid residues (positions 87 to 102) by using 24 pre-S deletion mutants; this binding domain coincides with a major virus-neutralizing antibody epitope. Furthermore, site-directed mutagenesis revealed that an arginine residue at position 97 is critical for p170 binding. p170 was purified by a combination of ion-exchange and affinity chromatographies, and four peptide sequences were obtained. Two peptides showed significant similarities to human and animal carboxypeptides H, M, and N. Taken together, these results raise the possibility that the p170 binding protein is important during the replication cycle of DHBV.

Full Text

The Full Text of this article is available as a PDF (514.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Budkowska A., Bedossa P., Groh F., Louise A., Pillot J. Fibronectin of human liver sinusoids binds hepatitis B virus: identification by an anti-idiotypic antibody bearing the internal image of the pre-S2 domain. J Virol. 1995 Feb;69(2):840–848. doi: 10.1128/jvi.69.2.840-848.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Budkowska A., Quan C., Groh F., Bedossa P., Dubreuil P., Bouvet J. P., Pillot J. Hepatitis B virus (HBV) binding factor in human serum: candidate for a soluble form of hepatocyte HBV receptor. J Virol. 1993 Jul;67(7):4316–4322. doi: 10.1128/jvi.67.7.4316-4322.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chassot S., Lambert V., Kay A., Godinot C., Roux B., Trepo C., Cova L. Fine mapping of neutralization epitopes on duck hepatitis B virus (DHBV) pre-S protein using monoclonal antibodies and overlapping peptides. Virology. 1993 Jan;192(1):217–223. doi: 10.1006/viro.1993.1024. [DOI] [PubMed] [Google Scholar]
  4. Cheung R. C., Robinson W. S., Marion P. L., Greenberg H. B. Epitope mapping of neutralizing monoclonal antibodies against duck hepatitis B virus. J Virol. 1989 Jun;63(6):2445–2451. doi: 10.1128/jvi.63.6.2445-2451.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fernholz D., Wildner G., Will H. Minor envelope proteins of duck hepatitis B virus are initiated at internal pre-S AUG codons but are not essential for infectivity. Virology. 1993 Nov;197(1):64–73. doi: 10.1006/viro.1993.1567. [DOI] [PubMed] [Google Scholar]
  6. Fricker L. D., Evans C. J., Esch F. S., Herbert E. Cloning and sequence analysis of cDNA for bovine carboxypeptidase E. Nature. 1986 Oct 2;323(6087):461–464. doi: 10.1038/323461a0. [DOI] [PubMed] [Google Scholar]
  7. Gebhard W., Schube M., Eulitz M. cDNA cloning and complete primary structure of the small, active subunit of human carboxypeptidase N (kininase 1). Eur J Biochem. 1989 Jan 2;178(3):603–607. doi: 10.1111/j.1432-1033.1989.tb14488.x. [DOI] [PubMed] [Google Scholar]
  8. Grgacic E. V., Anderson D. A. The large surface protein of duck hepatitis B virus is phosphorylated in the pre-S domain. J Virol. 1994 Nov;68(11):7344–7350. doi: 10.1128/jvi.68.11.7344-7350.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Halpern M. S., England J. M., Deery D. T., Petcu D. J., Mason W. S., Molnar-Kimber K. L. Viral nucleic acid synthesis and antigen accumulation in pancreas and kidney of Pekin ducks infected with duck hepatitis B virus. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4865–4869. doi: 10.1073/pnas.80.15.4865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hertogs K., Leenders W. P., Depla E., De Bruin W. C., Meheus L., Raymackers J., Moshage H., Yap S. H. Endonexin II, present on human liver plasma membranes, is a specific binding protein of small hepatitis B virus (HBV) envelope protein. Virology. 1993 Dec;197(2):549–557. doi: 10.1006/viro.1993.1628. [DOI] [PubMed] [Google Scholar]
  11. Ishikawa T., Kuroki K., Lenhoff R., Summers J., Ganem D. Analysis of the binding of a host cell surface glycoprotein to the preS protein of duck hepatitis B virus. Virology. 1994 Aug 1;202(2):1061–1064. doi: 10.1006/viro.1994.1440. [DOI] [PubMed] [Google Scholar]
  12. Klingmüller U., Schaller H. Hepadnavirus infection requires interaction between the viral pre-S domain and a specific hepatocellular receptor. J Virol. 1993 Dec;67(12):7414–7422. doi: 10.1128/jvi.67.12.7414-7422.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kuroki K., Cheung R., Marion P. L., Ganem D. A cell surface protein that binds avian hepatitis B virus particles. J Virol. 1994 Apr;68(4):2091–2096. doi: 10.1128/jvi.68.4.2091-2096.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kuroki K., Eng F., Ishikawa T., Turck C., Harada F., Ganem D. gp180, a host cell glycoprotein that binds duck hepatitis B virus particles, is encoded by a member of the carboxypeptidase gene family. J Biol Chem. 1995 Jun 23;270(25):15022–15028. doi: 10.1074/jbc.270.25.15022. [DOI] [PubMed] [Google Scholar]
  15. Lambert V., Fernholz D., Sprengel R., Fourel I., Deléage G., Wildner G., Peyret C., Trépo C., Cova L., Will H. Virus-neutralizing monoclonal antibody to a conserved epitope on the duck hepatitis B virus pre-S protein. J Virol. 1990 Mar;64(3):1290–1297. doi: 10.1128/jvi.64.3.1290-1297.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lenhoff R. J., Summers J. Coordinate regulation of replication and virus assembly by the large envelope protein of an avian hepadnavirus. J Virol. 1994 Jul;68(7):4565–4571. doi: 10.1128/jvi.68.7.4565-4571.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Li J. S., Cova L., Buckland R., Lambert V., Deléage G., Trépo C. Duck hepatitis B virus can tolerate insertion, deletion, and partial frameshift mutation in the distal pre-S region. J Virol. 1989 Nov;63(11):4965–4968. doi: 10.1128/jvi.63.11.4965-4968.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Macrae D. R., Bruss V., Ganem D. Myristylation of a duck hepatitis B virus envelope protein is essential for infectivity but not for virus assembly. Virology. 1991 Mar;181(1):359–363. doi: 10.1016/0042-6822(91)90503-4. [DOI] [PubMed] [Google Scholar]
  19. Mandart E., Kay A., Galibert F. Nucleotide sequence of a cloned duck hepatitis B virus genome: comparison with woodchuck and human hepatitis B virus sequences. J Virol. 1984 Mar;49(3):782–792. doi: 10.1128/jvi.49.3.782-792.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Manser E., Fernandez D., Loo L., Goh P. Y., Monfries C., Hall C., Lim L. Human carboxypeptidase E. Isolation and characterization of the cDNA, sequence conservation, expression and processing in vitro. Biochem J. 1990 Apr 15;267(2):517–525. doi: 10.1042/bj2670517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mehdi H., Kaplan M. J., Anlar F. Y., Yang X., Bayer R., Sutherland K., Peeples M. E. Hepatitis B virus surface antigen binds to apolipoprotein H. J Virol. 1994 Apr;68(4):2415–2424. doi: 10.1128/jvi.68.4.2415-2424.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Neurath A. R., Strick N., Li Y. Y. Cells transfected with human interleukin 6 cDNA acquire binding sites for the hepatitis B virus envelope protein. J Exp Med. 1992 Dec 1;176(6):1561–1569. doi: 10.1084/jem.176.6.1561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Persing D. H., Varmus H. E., Ganem D. The preS1 protein of hepatitis B virus is acylated at its amino terminus with myristic acid. J Virol. 1987 May;61(5):1672–1677. doi: 10.1128/jvi.61.5.1672-1677.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pontisso P., Ruvoletto M. G., Tiribelli C., Gerlich W. H., Ruol A., Alberti A. The preS1 domain of hepatitis B virus and IgA cross-react in their binding to the hepatocyte surface. J Gen Virol. 1992 Aug;73(Pt 8):2041–2045. doi: 10.1099/0022-1317-73-8-2041. [DOI] [PubMed] [Google Scholar]
  25. Pugh J. C., Simmons H. Duck hepatitis B virus infection of Muscovy duck hepatocytes and nature of virus resistance in vivo. J Virol. 1994 Apr;68(4):2487–2494. doi: 10.1128/jvi.68.4.2487-2494.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pugh J. C., Summers J. W. Infection and uptake of duck hepatitis B virus by duck hepatocytes maintained in the presence of dimethyl sulfoxide. Virology. 1989 Oct;172(2):564–572. doi: 10.1016/0042-6822(89)90199-2. [DOI] [PubMed] [Google Scholar]
  27. Sprengel R., Kaleta E. F., Will H. Isolation and characterization of a hepatitis B virus endemic in herons. J Virol. 1988 Oct;62(10):3832–3839. doi: 10.1128/jvi.62.10.3832-3839.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Summers J., Smith P. M., Huang M. J., Yu M. S. Morphogenetic and regulatory effects of mutations in the envelope proteins of an avian hepadnavirus. J Virol. 1991 Mar;65(3):1310–1317. doi: 10.1128/jvi.65.3.1310-1317.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sureau C., Guerra B., Lanford R. E. Role of the large hepatitis B virus envelope protein in infectivity of the hepatitis delta virion. J Virol. 1993 Jan;67(1):366–372. doi: 10.1128/jvi.67.1.366-372.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tagawa M., Omata M., Yokosuka O., Uchiumi K., Imazeki F., Okuda K. Early events in duck hepatitis B virus infection. Sequential appearance of viral deoxyribonucleic acid in the liver, pancreas, kidney, and spleen. Gastroenterology. 1985 Dec;89(6):1224–1229. [PubMed] [Google Scholar]
  31. Tan F., Chan S. J., Steiner D. F., Schilling J. W., Skidgel R. A. Molecular cloning and sequencing of the cDNA for human membrane-bound carboxypeptidase M. Comparison with carboxypeptidases A, B, H, and N. J Biol Chem. 1989 Aug 5;264(22):13165–13170. [PubMed] [Google Scholar]
  32. Tong S. P., Li J. S., Vitvitski L., Trépo C. Active hepatitis B virus replication in the presence of anti-HBe is associated with viral variants containing an inactive pre-C region. Virology. 1990 Jun;176(2):596–603. doi: 10.1016/0042-6822(90)90030-u. [DOI] [PubMed] [Google Scholar]
  33. Tuttleman J. S., Pugh J. C., Summers J. W. In vitro experimental infection of primary duck hepatocyte cultures with duck hepatitis B virus. J Virol. 1986 Apr;58(1):17–25. doi: 10.1128/jvi.58.1.17-25.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Yuasa S., Cheung R. C., Pham Q., Robinson W. S., Marion P. L. Peptide mapping of neutralizing and nonneutralizing epitopes of duck hepatitis B virus pre-S polypeptide. Virology. 1991 Mar;181(1):14–21. doi: 10.1016/0042-6822(91)90465-n. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES