Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1996 Jul;70(7):4567–4575. doi: 10.1128/jvi.70.7.4567-4575.1996

Covalently closed circular viral DNA formed from two types of linear DNA in woodchuck hepatitis virus-infected liver.

W Yang 1, W S Mason 1, J Summers 1
PMCID: PMC190393  PMID: 8676483

Abstract

We found that livers from woodchucks chronically infected with woodchuck hepatitis virus (WHV) contained covalently closed circular DNA (cccDNA) molecules with deletions and insertions indicative of their formation from linear viral DNA by nonhomologous recombination, as we previously described for the duck hepatitis B virus (W. Yang and J. Summers, J. Virol. 69:4029-4036, 1995). However, evidence for two different types of linear precursors was obtained by analysis of the recombination joints in WHV cccDNA. Type 1 linear precursors possessed the structural properties that correspond to those of in situ-primed linear DNA molecules, which constitute between 7 and 20% of all viral DNA replicative intermediates synthesized in the liver. Type 2 linear precursors are hypothetical species of linear DNAs with a terminal duplication of the cohesive-end region, between DR1 and DR2. This type of linear DNA has not been previously described and was not detected among the DNA species present in nucleocapsids. A fraction of cccDNAs formed from both type 1 and type 2 linear DNAs are predicted to be functional for further DNA synthesis, and some evidence for the formation of two or more generations of cccDNA from linear DNA was observed.

Full Text

The Full Text of this article is available as a PDF (365.7 KB).

Selected References

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

  1. Bartenschlager R., Junker-Niepmann M., Schaller H. The P gene product of hepatitis B virus is required as a structural component for genomic RNA encapsidation. J Virol. 1990 Nov;64(11):5324–5332. doi: 10.1128/jvi.64.11.5324-5332.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Condreay L. D., Wu T. T., Aldrich C. E., Delaney M. A., Summers J., Seeger C., Mason W. S. Replication of DHBV genomes with mutations at the sites of initiation of minus- and plus-strand DNA synthesis. Virology. 1992 May;188(1):208–216. doi: 10.1016/0042-6822(92)90751-a. [DOI] [PubMed] [Google Scholar]
  4. Enders G. H., Ganem D., Varmus H. E. 5'-terminal sequences influence the segregation of ground squirrel hepatitis virus RNAs into polyribosomes and viral core particles. J Virol. 1987 Jan;61(1):35–41. doi: 10.1128/jvi.61.1.35-41.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Feitelson M. A., Duan L. X., Guo J., Blumberg B. S. X region deletion mutants associated with surface antigen-positive hepatitis B virus infections. Gastroenterology. 1995 Jun;108(6):1810–1819. doi: 10.1016/0016-5085(95)90144-2. [DOI] [PubMed] [Google Scholar]
  6. Feitelson M. A., Duan L. X., Guo J., Sun B., Woo J., Steensma K., Horiike N., Blumberg B. S. X region deletion variants of hepatitis B virus in surface antigen-negative infections and non-A, non-B hepatitis. J Infect Dis. 1995 Sep;172(3):713–722. doi: 10.1093/infdis/172.3.713. [DOI] [PubMed] [Google Scholar]
  7. Feitelson M., Duan L. X., Horiike N., Clayton M. Hepatitis B X open reading frame deletion mutants isolated from atypical hepatitis B virus infections. J Hepatol. 1991;13 (Suppl 4):S58–S60. doi: 10.1016/0168-8278(91)90025-7. [DOI] [PubMed] [Google Scholar]
  8. Feitelson M., Lega L., Guo J., Resti M., Rossi M. E., Azzari C., Blumberg B. S., Vierucci A. Pathogenesis of posttransfusion viral hepatitis in children with beta-thalassemia. Hepatology. 1994 Mar;19(3):558–568. doi: 10.1002/hep.1840190304. [DOI] [PubMed] [Google Scholar]
  9. Horwich A. L., Furtak K., Pugh J., Summers J. Synthesis of hepadnavirus particles that contain replication-defective duck hepatitis B virus genomes in cultured HuH7 cells. J Virol. 1990 Feb;64(2):642–650. doi: 10.1128/jvi.64.2.642-650.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Junker-Niepmann M., Bartenschlager R., Schaller H. A short cis-acting sequence is required for hepatitis B virus pregenome encapsidation and sufficient for packaging of foreign RNA. EMBO J. 1990 Oct;9(10):3389–3396. doi: 10.1002/j.1460-2075.1990.tb07540.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kajino K., Jilbert A. R., Saputelli J., Aldrich C. E., Cullen J., Mason W. S. Woodchuck hepatitis virus infections: very rapid recovery after a prolonged viremia and infection of virtually every hepatocyte. J Virol. 1994 Sep;68(9):5792–5803. doi: 10.1128/jvi.68.9.5792-5803.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kew M. C., Miller R. H., Chen H. S., Tennant B. C., Purcell R. H. Mutant woodchuck hepatitis virus genomes from virions resemble rearranged hepadnaviral integrants in hepatocellular carcinoma. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10211–10215. doi: 10.1073/pnas.90.21.10211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kodama K., Ogasawara N., Yoshikawa H., Murakami S. Nucleotide sequence of a cloned woodchuck hepatitis virus genome: evolutional relationship between hepadnaviruses. J Virol. 1985 Dec;56(3):978–986. doi: 10.1128/jvi.56.3.978-986.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lien J. M., Aldrich C. E., Mason W. S. Evidence that a capped oligoribonucleotide is the primer for duck hepatitis B virus plus-strand DNA synthesis. J Virol. 1986 Jan;57(1):229–236. doi: 10.1128/jvi.57.1.229-236.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lien J. M., Petcu D. J., Aldrich C. E., Mason W. S. Initiation and termination of duck hepatitis B virus DNA synthesis during virus maturation. J Virol. 1987 Dec;61(12):3832–3840. doi: 10.1128/jvi.61.12.3832-3840.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Loeb D. D., Hirsch R. C., Ganem D. Sequence-independent RNA cleavages generate the primers for plus strand DNA synthesis in hepatitis B viruses: implications for other reverse transcribing elements. EMBO J. 1991 Nov;10(11):3533–3540. doi: 10.1002/j.1460-2075.1991.tb04917.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mason W. S., Aldrich C., Summers J., Taylor J. M. Asymmetric replication of duck hepatitis B virus DNA in liver cells: Free minus-strand DNA. Proc Natl Acad Sci U S A. 1982 Jul;79(13):3997–4001. doi: 10.1073/pnas.79.13.3997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mason W. S., Seal G., Summers J. Virus of Pekin ducks with structural and biological relatedness to human hepatitis B virus. J Virol. 1980 Dec;36(3):829–836. doi: 10.1128/jvi.36.3.829-836.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nassal M., Junker-Niepmann M., Schaller H. Translational inactivation of RNA function: discrimination against a subset of genomic transcripts during HBV nucleocapsid assembly. Cell. 1990 Dec 21;63(6):1357–1363. doi: 10.1016/0092-8674(90)90431-d. [DOI] [PubMed] [Google Scholar]
  20. Seeger C., Ganem D., Varmus H. E. Biochemical and genetic evidence for the hepatitis B virus replication strategy. Science. 1986 Apr 25;232(4749):477–484. doi: 10.1126/science.3961490. [DOI] [PubMed] [Google Scholar]
  21. Seeger C., Maragos J. Identification and characterization of the woodchuck hepatitis virus origin of DNA replication. J Virol. 1990 Jan;64(1):16–23. doi: 10.1128/jvi.64.1.16-23.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Seeger C., Maragos J. Molecular analysis of the function of direct repeats and a polypurine tract for plus-strand DNA priming in woodchuck hepatitis virus. J Virol. 1989 May;63(5):1907–1915. doi: 10.1128/jvi.63.5.1907-1915.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Staprans S., Loeb D. D., Ganem D. Mutations affecting hepadnavirus plus-strand DNA synthesis dissociate primer cleavage from translocation and reveal the origin of linear viral DNA. J Virol. 1991 Mar;65(3):1255–1262. doi: 10.1128/jvi.65.3.1255-1262.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Summers J., Mason W. S. Replication of the genome of a hepatitis B--like virus by reverse transcription of an RNA intermediate. Cell. 1982 Jun;29(2):403–415. doi: 10.1016/0092-8674(82)90157-x. [DOI] [PubMed] [Google Scholar]
  25. Summers J., O'Connell A., Millman I. Genome of hepatitis B virus: restriction enzyme cleavage and structure of DNA extracted from Dane particles. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4597–4601. doi: 10.1073/pnas.72.11.4597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Summers J., Smith P. M., Horwich A. L. Hepadnavirus envelope proteins regulate covalently closed circular DNA amplification. J Virol. 1990 Jun;64(6):2819–2824. doi: 10.1128/jvi.64.6.2819-2824.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Summers J., Smolec J. M., Snyder R. A virus similar to human hepatitis B virus associated with hepatitis and hepatoma in woodchucks. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4533–4537. doi: 10.1073/pnas.75.9.4533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tuttleman J. S., Pourcel C., Summers J. Formation of the pool of covalently closed circular viral DNA in hepadnavirus-infected cells. Cell. 1986 Nov 7;47(3):451–460. doi: 10.1016/0092-8674(86)90602-1. [DOI] [PubMed] [Google Scholar]
  30. Wang G. H., Seeger C. Novel mechanism for reverse transcription in hepatitis B viruses. J Virol. 1993 Nov;67(11):6507–6512. doi: 10.1128/jvi.67.11.6507-6512.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wang H. P., Rogler C. E. Topoisomerase I-mediated integration of hepadnavirus DNA in vitro. J Virol. 1991 May;65(5):2381–2392. doi: 10.1128/jvi.65.5.2381-2392.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wu T. T., Coates L., Aldrich C. E., Summers J., Mason W. S. In hepatocytes infected with duck hepatitis B virus, the template for viral RNA synthesis is amplified by an intracellular pathway. Virology. 1990 Mar;175(1):255–261. doi: 10.1016/0042-6822(90)90206-7. [DOI] [PubMed] [Google Scholar]
  33. Yang W., Summers J. Illegitimate replication of linear hepadnavirus DNA through nonhomologous recombination. J Virol. 1995 Jul;69(7):4029–4036. doi: 10.1128/jvi.69.7.4029-4036.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES