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. 1993 Jan;67(1):446–454. doi: 10.1128/jvi.67.1.446-454.1993

Hepatitis delta virus antigen forms dimers and multimeric complexes in vivo.

J G Wang 1, S M Lemon 1
PMCID: PMC237381  PMID: 7677957

Abstract

Although the hepatitis delta virus genome contains multiple open reading frames, only one of these reading frames is known to be expressed during replication of the virus. This open reading frame encodes two distinct molecular species of hepatitis delta antigen (HDAg), p24 delta and p27 delta, depending on the location of the stop codon which terminates translation. We found antibody specific for p27 delta to be capable of precipitating p24 delta in extracts of infected liver, indicating that p27 delta and p24 delta form heterologous complexes in vivo. After cross-linking with 0.05% glutaraldehyde, specific HDAg dimers were detected in antigen prepared from both the liver and serum of an HDV-infected woodchuck carrier of woodchuck hepatitis virus. Guanidine HCl-denatured HDAg extracted from liver and dialyzed against phosphate-buffered saline sedimented in rate-zonal sucrose density gradients as 15S multimeric complexes. These 15S multimers were stable in the presence of 1.2% Nonidet P-40. After RNase digestion, the 15S complex was reduced to a 12S complex without associated RNA, while boiling for 3 min in 1% sodium dodecyl sulfate-0.5% 2-mercaptoethanol further reduced the 15S complex to 3S HDAg monomers. In the absence of glutaraldehyde cross-linking, HDAg extracted from liver migrated as monomer species in reducing and nonreducing gels, suggesting that the conserved cysteine residue present in p27 delta does not play a role in the formation of either dimers or multimers. On the other hand, an amino-terminal chymotrypsin-digested HDAg fragment, with a predicted length of 81 or less amino acids, retained the ability to form dimers, consistent with the hypothesis that a coiled-coil motif present between residues 27 and 58 may play a role in HDAg protein interactions in vivo.

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Selected References

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

  1. Bergmann K. F., Cote P. J., Moriarty A., Gerin J. L. Hepatitis delta antigen. Antigenic structure and humoral immune response. J Immunol. 1989 Dec 1;143(11):3714–3721. [PubMed] [Google Scholar]
  2. Bergmann K. F., Gerin J. L. Antigens of hepatitis delta virus in the liver and serum of humans and animals. J Infect Dis. 1986 Oct;154(4):702–706. doi: 10.1093/infdis/154.4.702. [DOI] [PubMed] [Google Scholar]
  3. Chang F. L., Chen P. J., Tu S. J., Wang C. J., Chen D. S. The large form of hepatitis delta antigen is crucial for assembly of hepatitis delta virus. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8490–8494. doi: 10.1073/pnas.88.19.8490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chang M. F., Baker S. C., Soe L. H., Kamahora T., Keck J. G., Makino S., Govindarajan S., Lai M. M. Human hepatitis delta antigen is a nuclear phosphoprotein with RNA-binding activity. J Virol. 1988 Jul;62(7):2403–2410. doi: 10.1128/jvi.62.7.2403-2410.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chao M., Hsieh S. Y., Taylor J. Role of two forms of hepatitis delta virus antigen: evidence for a mechanism of self-limiting genome replication. J Virol. 1990 Oct;64(10):5066–5069. doi: 10.1128/jvi.64.10.5066-5069.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Glenn J. S., Watson J. A., Havel C. M., White J. M. Identification of a prenylation site in delta virus large antigen. Science. 1992 May 29;256(5061):1331–1333. doi: 10.1126/science.1598578. [DOI] [PubMed] [Google Scholar]
  7. Kuo M. Y., Goldberg J., Coates L., Mason W., Gerin J., Taylor J. Molecular cloning of hepatitis delta virus RNA from an infected woodchuck liver: sequence, structure, and applications. J Virol. 1988 Jun;62(6):1855–1861. doi: 10.1128/jvi.62.6.1855-1861.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lai M. M., Chao Y. C., Chang M. F., Lin J. H., Gust I. Functional studies of hepatitis delta antigen and delta virus RNA. Prog Clin Biol Res. 1991;364:283–292. [PubMed] [Google Scholar]
  9. Lin J. H., Chang M. F., Baker S. C., Govindarajan S., Lai M. M. Characterization of hepatitis delta antigen: specific binding to hepatitis delta virus RNA. J Virol. 1990 Sep;64(9):4051–4058. doi: 10.1128/jvi.64.9.4051-4058.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Luo G. X., Chao M., Hsieh S. Y., Sureau C., Nishikura K., Taylor J. A specific base transition occurs on replicating hepatitis delta virus RNA. J Virol. 1990 Mar;64(3):1021–1027. doi: 10.1128/jvi.64.3.1021-1027.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Macnaughton T. B., Gowans E. J., Reinboth B., Jilbert A. R., Burrell C. J. Stable expression of hepatitis delta virus antigen in a eukaryotic cell line. J Gen Virol. 1990 Jun;71(Pt 6):1339–1345. doi: 10.1099/0022-1317-71-6-1339. [DOI] [PubMed] [Google Scholar]
  12. Makino S., Chang M. F., Shieh C. K., Kamahora T., Vannier D. M., Govindarajan S., Lai M. M. Molecular cloning and sequencing of a human hepatitis delta (delta) virus RNA. Nature. 1987 Sep 24;329(6137):343–346. doi: 10.1038/329343a0. [DOI] [PubMed] [Google Scholar]
  13. Negro F., Bergmann K. F., Baroudy B. M., Satterfield W. C., Popper H., Purcell R. H., Gerin J. L. Chronic hepatitis D virus (HDV) infection in hepatitis B virus carrier chimpanzees experimentally superinfected with HDV. J Infect Dis. 1988 Jul;158(1):151–159. doi: 10.1093/infdis/158.1.151. [DOI] [PubMed] [Google Scholar]
  14. Negro F., Korba B. E., Forzani B., Baroudy B. M., Brown T. L., Gerin J. L., Ponzetto A. Hepatitis delta virus (HDV) and woodchuck hepatitis virus (WHV) nucleic acids in tissues of HDV-infected chronic WHV carrier woodchucks. J Virol. 1989 Apr;63(4):1612–1618. doi: 10.1128/jvi.63.4.1612-1618.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nibert M. L., Dermody T. S., Fields B. N. Structure of the reovirus cell-attachment protein: a model for the domain organization of sigma 1. J Virol. 1990 Jun;64(6):2976–2989. doi: 10.1128/jvi.64.6.2976-2989.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. O'Shea E. K., Klemm J. D., Kim P. S., Alber T. X-ray structure of the GCN4 leucine zipper, a two-stranded, parallel coiled coil. Science. 1991 Oct 25;254(5031):539–544. doi: 10.1126/science.1948029. [DOI] [PubMed] [Google Scholar]
  17. Ponzetto A., Cote P. J., Popper H., Hoyer B. H., London W. T., Ford E. C., Bonino F., Purcell R. H., Gerin J. L. Transmission of the hepatitis B virus-associated delta agent to the eastern woodchuck. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2208–2212. doi: 10.1073/pnas.81.7.2208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rizzetto M., Hoyer B., Canese M. G., Shih J. W., Purcell R. H., Gerin J. L. delta Agent: association of delta antigen with hepatitis B surface antigen and RNA in serum of delta-infected chimpanzees. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6124–6128. doi: 10.1073/pnas.77.10.6124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rizzetto M., Shih J. W., Gerin J. L. The hepatitis B virus-associated delta antigen: isolation from liver, development of solid-phase radioimmunoassays for delta antigen and anti-delta and partial characterization of delta antigen. J Immunol. 1980 Jul;125(1):318–324. [PubMed] [Google Scholar]
  20. Ryu W. S., Bayer M., Taylor J. Assembly of hepatitis delta virus particles. J Virol. 1992 Apr;66(4):2310–2315. doi: 10.1128/jvi.66.4.2310-2315.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Taylor J. M. Hepatitis delta virus: cis and trans functions required for replication. Cell. 1990 May 4;61(3):371–373. doi: 10.1016/0092-8674(90)90516-h. [DOI] [PubMed] [Google Scholar]
  22. Wang J. G., Cullen J., Lemon S. M. Immunoblot analysis demonstrates that the large and small forms of hepatitis delta virus antigen have different C-terminal amino acid sequences. J Gen Virol. 1992 Jan;73(Pt 1):183–188. doi: 10.1099/0022-1317-73-1-183. [DOI] [PubMed] [Google Scholar]
  23. Wang J. G., Jansen R. W., Brown E. A., Lemon S. M. Immunogenic domains of hepatitis delta virus antigen: peptide mapping of epitopes recognized by human and woodchuck antibodies. J Virol. 1990 Mar;64(3):1108–1116. doi: 10.1128/jvi.64.3.1108-1116.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wang K. S., Choo Q. L., Weiner A. J., Ou J. H., Najarian R. C., Thayer R. M., Mullenbach G. T., Denniston K. J., Gerin J. L., Houghton M. Structure, sequence and expression of the hepatitis delta (delta) viral genome. Nature. 1986 Oct 9;323(6088):508–514. doi: 10.1038/323508a0. [DOI] [PubMed] [Google Scholar]
  25. Weiner A. J., Choo Q. L., Wang K. S., Govindarajan S., Redeker A. G., Gerin J. L., Houghton M. A single antigenomic open reading frame of the hepatitis delta virus encodes the epitope(s) of both hepatitis delta antigen polypeptides p24 delta and p27 delta. J Virol. 1988 Feb;62(2):594–599. doi: 10.1128/jvi.62.2.594-599.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Xia Y. P., Yeh C. T., Ou J. H., Lai M. M. Characterization of nuclear targeting signal of hepatitis delta antigen: nuclear transport as a protein complex. J Virol. 1992 Feb;66(2):914–921. doi: 10.1128/jvi.66.2.914-921.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

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