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. 2002 Feb;76(4):1986-90.
doi: 10.1128/jvi.76.4.1986-1990.2002.

Inhibition of duck hepatitis B virus infection by a myristoylated pre-S peptide of the large viral surface protein

Stephan Urban  1 , Philippe Gripon
Affiliations

Inhibition of duck hepatitis B virus infection by a myristoylated pre-S peptide of the large viral surface protein

Stephan Urban et al. J Virol. 2002 Feb.

Abstract

We have used the duck hepatitis B virus (DHBV) model to study the interference with infection by a myristoylated peptide representing an N-terminal pre-S subdomain of the large viral envelope protein. Although lacking the essential part of the carboxypeptidase D (formerly called gp180) receptor binding site, the peptide binds hepatocytes and subsequently blocks DHBV infection. Since its activity requires an amino acid sequence involved in host discrimination between DHBV and the related heron HBV (T. Ishikawa and D. Ganem, Proc. Natl. Acad. Sci. USA 92:6259-6263, 1995), we suggest that it is related to the postulated host-discriminating cofactor of infection.

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Figures

FIG. 1.
FIG. 1.
Schematic illustration of the pre-S domain of the DHBV L protein and the two DHBV pre-S-derived myristoylated peptides Dpre-S2-41myr and Dpre-S2-21myr (A) and the primary sequences of myristoylated Dpre-S2-41myr, Hepre-S2-44myr, and Hupre-S2-48myr (B) used in DHBV infection competition experiments. (A) The duCPD (gp180) receptor binding site (amino acids 30 to 115) within the pre-S domain (amino acids 1 to 161) of the DHBV L protein consists of an essential, partially α-helical subdomain (amino acids 86 to 115) and a most-randomly structured element (amino acids 30 to 85) which is needed for the formation of a high-affinity complex. Directly adjacent to the receptor binding site is Ser-118, which becomes phosphorylated (13). Partially overlapping with the stabilizing element but clearly extending it at its N terminus is the sequence that recovers infectivity of HHBV for PDHs (amino acids 22 to 37). (B) Sequence alignment of the two inhibitory peptides Dpre-S2-41myr and the homologous Hepre-S2-44myr. Sequence identity is 57%. Note that within the host-determining region (boxed sequence between amino acids 22 and 37), HHBV pre-S contains the additional insertion 31-PEF-33. The HBV pre-S-derived peptide Hupre-S2-48myr displays no significant homology to its avian counterparts.
FIG. 2.
FIG. 2.
Competition of DHBV infection by the myristoylated pre-S-derived peptides Dpre-S2-41myr, Dpre-S2-21myr, Hepre-S2-44myr, and Hupre-S2-48myr. PDHs (8 × 105) were infected at a multiplicity of infection of ≈100 in the presence of 5, 20, 50, 100, and 400 nM concentrations of the respective pre-S peptides. Fourteen hours later, peptides and virus were removed. (A) At 7 days postinfection total intracellular DNA was extracted and analyzed by dot blotting as described previously (12). (B) Viral DNA was quantified with a PhosphorImager, and results are presented as percentages of the value for an uncompeted control infection In each group of four bars, results for Dpre-S2-41myr, Dpre-S2-21myr, Hepre-S2-44myr, and Hupre-S2-48myr are shown from left to right, respectively. Three independent experiments were performed, and results from one representative experiment are shown.
FIG. 3.
FIG. 3.
Competition of DHBV infection by the nonmyristoylated pre-S-derived peptides Dpre-S1-41, Dpre-S1-21, Hepre-S1-44, and Hupre-S1-48. PDHs (8 × 105) were infected overnight at a multiplicity of infection of ≈100 in the presence of 0.2, 1, 5, 20, and 100 μM concentrations of the respective pre-S peptides. Fourteen hours later, peptides and virus were removed. (A) At 7 days postinfection total intracellular DNA was extracted and analyzed by dot blot analysis as described previously (12). (B) Viral DNA was quantified with a PhosphorImager, and results are presented as percentages of the value for an uncompeted control infection. In each group of four bars, results for Dpre-S1-41, Dpre-S1-21, Hepre-S1-44, and Hupre-S1-48 are shown from left to right, respectively. Two independent experiments were performed.
FIG. 4.
FIG. 4.
(A) Competition of DHBV infection following preincubation of PDHs with Dpre-S2-41myr. PDHs were incubated with Dpre-S2-48myr (800 nM) for 15, 30, 90, 180, and 360 min prior to infection with DHBV (14 h; multiplicity of infection, ≈75). Seven days later intracellular viral DNA was extracted, analyzed by dot blotting. and quantified with a PhosphorImager. Values are given as percentages of the value for an uncompeted infection. Error bars indicate standard deviations. (B) Association of Dpre-S1-41 with PDHs. NHS-rhodamine (Pierce) was chemically coupled to Dpre-S1-41 according to the manufacturer's protocol. After purification on a PD10 gel filtration column (Amersham-Pharmacia), the labeled peptide was incubated with PDHs for 3 h. After removal of surplus free peptide, cell-associated Dpre-S1-41 was visualized by fluorescence microscopy (red). The phase-contrast image is shown in blue.
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References

    1. Borst, E. M. 1997. Untersuchungen zur Interaktion des Enten Hepatitis B Virus L-Proteins mit primären Entenhepatozyten. Ph.D. thesis. University of Ulm, Ulm, Germany.
    1. Breiner, K. M., S. Urban, and H. Schaller. 1998. Carboxypeptidase D (gp180), a Golgi-resident protein, functions in the attachment and entry of avian hepatitis B viruses. J. Virol. 72:8098-8104. - PMC - PubMed
    1. Chan, D. C., and P. S. Kim. 1998. HIV entry and its inhibition. Cell 93:681-684. - PubMed
    1. Gripon, P., J. Le Seyec, S. Rumin, and C. Guguen-Guillouzo. 1995. Myristylation of the hepatitis B virus large surface protein is essential for viral infectivity. Virology 213:292-299. - PubMed
    1. Ishikawa, T., K. Kuroki, R. Lenhoff, J. Summers, and D. Ganem. 1994. Analysis of the binding of a host cell surface glycoprotein to the pre-S protein of duck hepatitis B virus. Virology 202:1061-1064. - PubMed

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