Localization of Duck Hepatitis B Virus Polymerase Within Cells

  • Ermei Yao
  • John E. Tavis
Part of the Methods in Molecular Medicine book series (MIMM, volume 95)


Hepadnaviruses are small, DNA-containing viruses that replicate by reverse transcription (1). They have a lipid envelope surrounding an icosahedral protein core particle, whose shell is composed of a single viral protein, the core protein. Within the core particle, the viral reverse transcriptase (polymerase) is covalently linked to the double-stranded viral genome.


Western Analysis Core Particle Laemmli Buffer Antifade Agent Single Viral Protein 
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  1. 1.
    Ganem, D. and Schneider, R. J. (2001) Hepadnaviridae: the viruses and their replication. In: Knipe, D. M. Howley, P. M. Griffen, D. E. et al. (eds.), Fields Virology, 4th ed., vol 2, Lippencott Williams & Wilkins, Philadelphia, PA, 2923–2969.Google Scholar
  2. 2.
    Bartenschlager, R., Junker-Niepmann, M., and Schaller, H. (1990) The P gene product of hepatitis B virus is required as a structural component for genomic RNA encapsidation. J. Virol. 64, 5324–5332.PubMedGoogle Scholar
  3. 3.
    Junker-Niepmann, M., Bartenschlager, R., and Schaller, H. (1990) A short cis-acting sequence is required for hepatitis B virus pregenome encapsidation and sufficient for packaging of foreign RNA. EMBO J. 9, 3389–3396.PubMedGoogle Scholar
  4. 4.
    Hirsch, R. C., Lavine, J. E. Chang, L. J. Varmus, H. E. and Ganem, D. (1990) Polymerase gene products of hepatitis B viruses are required for genomic RNA packaging as well as for reverse transcription. Nature 344, 552–555.PubMedCrossRefGoogle Scholar
  5. 5.
    Tavis, J. E. and Ganem, D. (1996) Evidence for the activation of the hepatitis B virus polymerase by binding of its RNA template. J. Virol. 70, 5741–5750.PubMedGoogle Scholar
  6. 6.
    Tavis, J. E., Massey, B., and Gong, Y. (1998) The duck hepatitis B virus polymerase is activated by its RNA packaging signal, epsilon. J. Virol. 72, 5789–5796.PubMedGoogle Scholar
  7. 7.
    Wang, G.-H. and Seeger, C. (1993) Novel mechanism for reverse transcription in hepatitis B viruses. J. Virol. 67, 6507–6512.PubMedGoogle Scholar
  8. 8.
    Tavis, J. E., Perri, S., and Ganem, D. (1994) Hepadnavirus reverse transcription initiates within the stem-loop of the RNA packaging signal and employs a novel strand transfer. J. Virol. 68, 3536–3543.PubMedGoogle Scholar
  9. 9.
    Tavis, J. E. and Ganem, D. (1995) RNA sequences controlling the initiation and transfer of duck hepatitis B virus minus-strand DNA. J. Virol. 69, 4283–4291.PubMedGoogle Scholar
  10. 10.
    Nassal, M. and Rieger, A. (1996) A bulged region of the hepatitis B virus RNA encapsidation signal contains the replication origin for discontinuous first-strand DNA synthesis. J. Virol. 70, 2764–2773.PubMedGoogle Scholar
  11. 11.
    Chang, L.-J., Hirsch, R. C., Ganem, D., and Varmus, H. E. (1990) Effects of insertional and point mutations on the functions of the duck hepatitis B virus polymerase. J. Virol. 64, 5553–5558.PubMedGoogle Scholar
  12. 12.
    Radziwill, G., Tucker, W., and Schaller, H. (1990) Mutational analysis of the hepatitis B virus P gene product: domain structure and RNase H activity. J. Virol. 64, 613–620.PubMedGoogle Scholar
  13. 13.
    Zoulim, F. and Seeger, C. (1994) Reverse transcription in hepatitis B viruses is primed by a tyrosine residue of the polymerase. J. Virol. 68, 6–13.PubMedGoogle Scholar
  14. 14.
    Weber, M., Bronsema, V., Bartos, H., Bosserhoff, A., Bartenschlager, R., and Schaller, H. (1994) Hepadnavirus P protein utilizes a tyrosine residue in the TP domain to prime reverse transcription. J. Virol. 68, 2994–2999.PubMedGoogle Scholar
  15. 15.
    McClure, M. A. (1993) Evolutionary history of reverse transcriptase. In: Skalka, A. M. and Goff, S. P. (eds.), Reverse Transcriptase, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 425–444.Google Scholar
  16. 16.
    Poch, O., Sauvaget, I., Delarue, M., and Tordo, N. (1989) Identification of four conserved motifs among the RNA-dependent polymerase encoding elements. EMBO J. 8, 3867–3874.PubMedGoogle Scholar
  17. 17.
    Li, M. D., Bronson, D. L., Lemke, T. D., and Faras, A. J. (1995) Phylogenetic analyses of 55 retroelements on the basis of the nucleotide and product amino acid sequences of the pol gene. Mol. Biol. Evol. 12, 657–670.PubMedGoogle Scholar
  18. 18.
    Bavand, M., Feitelson, M., and Laub, O. (1989) The hepatitis B virus-associated reverse transcriptase is encoded by the viral pol gene. J. Virol. 63, 1019–1021.PubMedGoogle Scholar
  19. 19.
    Mack, D. H., Bloch, W., Nath, N., and Sninsky, J. J. (1988) Hepatitis B virus particles contain a polypeptide encoded by the largest open reading frame: a putative reverse transcriptas. J. Virol. 62, 4786–4790.PubMedGoogle Scholar
  20. 20.
    Bartenschlager, R. and Schaller, H. (1988) The amino-terminal domain of the hepadnaviral P-gene encodes the terminal protein (genome-linked protein) believed to prime reverse transcription. EMBO J. 7, 4185–4192.PubMedGoogle Scholar
  21. 21.
    Huang, H.-L., Jeng, K.-S., Hu, C.-P., Tsai, C.-H., Lo, S. J., and Chang, C. (2000) Identification and characterization of a structural protein of hepatitis B virus: a polymerase and surface fusion protein encoded by a spliced RNA. Virology 275, 398–410.PubMedCrossRefGoogle Scholar
  22. 22.
    Bartenschlager, R., Kuhn, C., and Schaller, H. (1992) Expression of the P-protein of the human hepatitis B virus in a vaccinia virus system and detection of the nucleocapsid-associated P-gene product by radiolabelling at newly introduced phosphorylation sites. Nucl. Acids Res. 20, 195–202.PubMedCrossRefGoogle Scholar
  23. 23.
    Oberhaus, S. M. and Newbold, J. E. (1996) Preparations of duck hepatitis B virions contain multiple DNA polymerase activities. Virology 226, 132–134.PubMedCrossRefGoogle Scholar
  24. 24.
    Oberhaus, S. M. and Newbold, J. E. (1993) Detection of DNA polymerase activities associated with purified duck hepatitis B virus core particles by using an activity gel assay. J. Virol. 67, 6558–6566.PubMedGoogle Scholar
  25. 25.
    Bavand, M. R. and Laub, O. (1988) Two proteins with reverse transcriptase activities associated with hepatitis B virus-like particles. J. Virol. 62, 626–628.PubMedGoogle Scholar
  26. 26.
    zu Putlitz, J., Lanford, R. E., Carlson, R. I., Notvall, L., De la Monte, S. M. and Wands, J. R. (1999) Properties of monoclonal antibodies directed against hepatitis B virus polymerase protein. J. Virol. 73, 4188–4196.PubMedGoogle Scholar
  27. 27.
    Yao, E., Gong, Y., Chen, N., and Tavis, J. E. (2000) The majority of duck hepatitis B virus reverse transcriptase in cells is nonencapsidated and is bound to a cytoplasmic structure. J. Virol. 74, 8648–8657.PubMedCrossRefGoogle Scholar
  28. 28.
    Radziwill, G., Zentgraf, H., Schaller, H., and Bosch, V. (1988) The duck hepatitis B virus DNA polymerase is tightly associated with the viral core structure and unable to switch to an exogenous template. Virology 163, 123–132.PubMedCrossRefGoogle Scholar
  29. 29.
    Sprengel, R., Kuhn, C., Will, H., and Schaller, H. (1985) Comparative sequence analysis of duck and human hepatitis B virus genomes. J. Med. Virol. 15, 323–333.PubMedCrossRefGoogle Scholar
  30. 30.
    Condreay, L. D., Aldrich, C. E., Coates, L., Mason, W. S., and Wu, T.-T. (1990) Efficient duck hepatitis B virus production by an avian liver tumor cell line. J. Virol. 64, 3249–3258.PubMedGoogle Scholar
  31. 31.
    Pollack, J. R. and Ganem, D. (1994) Site-specific RNA binding by a hepatitis B virus reverse transcriptase initiates two distinct reactions: RNA packaging and DNA synthesis. J. Virol. 68, 5579–5587.PubMedGoogle Scholar
  32. 32.
    Kozak, M. (1991) Structural features in eukaryotic mRNAs that modulate the initiation of translation. J. Biol. Chem. 266, 19,867–19,870.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  • Ermei Yao
    • 1
  • John E. Tavis
    • 1
  1. 1.Department of Molecular Microbiology and ImmunologySt. Louis University School of MedicineSt. Louis

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