Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1984 Sep;81(18):5849–5852. doi: 10.1073/pnas.81.18.5849

The cloned genome of ground squirrel hepatitis virus is infectious in the animal.

C Seeger, D Ganem, H E Varmus
PMCID: PMC391809  PMID: 6091114

Abstract

The lack of an in vitro infectivity assay for hepatitis B viruses has impeded the analysis of their genetic organization. To examine the feasibility of generating mutant and recombinant viruses after manipulation of cloned viral DNA in vitro, we have tested the infectivity of the cloned genome of ground squirrel hepatitis virus (GSHV) in virus-free Beechey ground squirrels. We demonstrate that cloned GSHV DNA is infectious when injected directly into the liver in the form of trimeric, head-to-tail recombinant clones and recircularized monomeric molecules but not when injected into the portal vein. Infections established in all four recipients of intrahepatic injections of cloned GSHV DNA exhibited the characteristics observed after administration of virus: GSHV surface antigen and viral DNA appeared in the serum 14-22 weeks after inoculation, and both circular and heterogeneous protein-linked forms of viral DNA were found in liver biopsy samples. Furthermore, virus present in the sera of these animals can be transmitted to other ground squirrels. These findings imply that any function of virion proteins in the initiation of infection by hepatitis B viruses can be bypassed with the use of cloned viral DNA and that this animal model is suitable for testing mutant genomes.

Full text

PDF
5849

Images in this article

5851Image
on p.5851
5851Image
on p.5851

Selected References

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

  1. 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]
  2. Cattaneo R., Will H., Hernandez N., Schaller H. Signals regulating hepatitis B surface antigen transcription. Nature. 1983 Sep 22;305(5932):336–338. doi: 10.1038/305336a0. [DOI] [PubMed] [Google Scholar]
  3. Feitelson M. A., Marion P. L., Robinson W. S. The nature of polypeptides larger in size than the major surface antigen components of hepatitis b and like viruses in ground squirrels, woodchucks, and ducks. Virology. 1983 Oct 15;130(1):76–90. doi: 10.1016/0042-6822(83)90119-8. [DOI] [PubMed] [Google Scholar]
  4. Galibert F., Chen T. N., Mandart E. Nucleotide sequence of a cloned woodchuck hepatitis virus genome: comparison with the hepatitis B virus sequence. J Virol. 1982 Jan;41(1):51–65. doi: 10.1128/jvi.41.1.51-65.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Galibert F., Mandart E., Fitoussi F., Tiollais P., Charnay P. Nucleotide sequence of the hepatitis B virus genome (subtype ayw) cloned in E. coli. Nature. 1979 Oct 25;281(5733):646–650. doi: 10.1038/281646a0. [DOI] [PubMed] [Google Scholar]
  6. Ganem D., Greenbaum L., Varmus H. E. Virion DNA of ground squirrel hepatitis virus: structural analysis and molecular cloning. J Virol. 1982 Oct;44(1):374–383. doi: 10.1128/jvi.44.1.374-383.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ganem D., Weiser B., Barchuk A., Brown R. J., Varmus H. E. Biological characterization of acute infection with ground squirrel hepatitis virus. J Virol. 1982 Oct;44(1):366–373. doi: 10.1128/jvi.44.1.366-373.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gerlich W. H., Robinson W. S. Hepatitis B virus contains protein attached to the 5' terminus of its complete DNA strand. Cell. 1980 Oct;21(3):801–809. doi: 10.1016/0092-8674(80)90443-2. [DOI] [PubMed] [Google Scholar]
  9. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  10. Kaplan P. M., Greenman R. L., Gerin J. L., Purcell R. H., Robinson W. S. DNA polymerase associated with human hepatitis B antigen. J Virol. 1973 Nov;12(5):995–1005. doi: 10.1128/jvi.12.5.995-1005.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Marion P. L., Oshiro L. S., Regnery D. C., Scullard G. H., Robinson W. S. A virus in Beechey ground squirrels that is related to hepatitis B virus of humans. Proc Natl Acad Sci U S A. 1980 May;77(5):2941–2945. doi: 10.1073/pnas.77.5.2941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mason W. S., Halpern M. S., England J. M., Seal G., Egan J., Coates L., Aldrich C., Summers J. Experimental transmission of duck hepatitis B virus. Virology. 1983 Dec;131(2):375–384. doi: 10.1016/0042-6822(83)90505-6. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Molnar-Kimber K. L., Summers J., Taylor J. M., Mason W. S. Protein covalently bound to minus-strand DNA intermediates of duck hepatitis B virus. J Virol. 1983 Jan;45(1):165–172. doi: 10.1128/jvi.45.1.165-172.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pasek M., Goto T., Gilbert W., Zink B., Schaller H., MacKay P., Leadbetter G., Murray K. Hepatitis B virus genes and their expression in E. coli. Nature. 1979 Dec 6;282(5739):575–579. doi: 10.1038/282575a0. [DOI] [PubMed] [Google Scholar]
  16. Ruiz-Opazo N., Chakraborty P. R., Shafritz D. A. Evidence for supercoiled hepatitis B virus DNA in chimpanzee liver and serum Dane particles: possible implications in persistent HBV infection. Cell. 1982 May;29(1):129–136. doi: 10.1016/0092-8674(82)90097-6. [DOI] [PubMed] [Google Scholar]
  17. Seeger C., Ganem D., Varmus H. E. Nucleotide sequence of an infectious molecularly cloned genome of ground squirrel hepatitis virus. J Virol. 1984 Aug;51(2):367–375. doi: 10.1128/jvi.51.2.367-375.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Siddiqui A., Marion P. L., Robinson W. S. Ground squirrel hepatitis virus DNA: molecular cloning and comparison with hepatitis B virus DNA. J Virol. 1981 Apr;38(1):393–397. doi: 10.1128/jvi.38.1.393-397.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Standring D. N., Rutter W. J., Varmus H. E., Ganem D. Transcription of the hepatitis B surface antigen gene in cultured murine cells initiates within the presurface region. J Virol. 1984 May;50(2):563–571. doi: 10.1128/jvi.50.2.563-571.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Stibbe W., Gerlich W. H. Structural relationships between minor and major proteins of hepatitis B surface antigen. J Virol. 1983 May;46(2):626–628. doi: 10.1128/jvi.46.2.626-628.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Summers J. Three recently described animal virus models for human hepatitis B virus. Hepatology. 1981 Mar-Apr;1(2):179–183. doi: 10.1002/hep.1840010215. [DOI] [PubMed] [Google Scholar]
  23. Tiollais P., Charnay P., Vyas G. N. Biology of hepatitis B virus. Science. 1981 Jul 24;213(4506):406–411. doi: 10.1126/science.6264599. [DOI] [PubMed] [Google Scholar]
  24. Weiser B., Ganem D., Seeger C., Varmus H. E. Closed circular viral DNA and asymmetrical heterogeneous forms in livers from animals infected with ground squirrel hepatitis virus. J Virol. 1983 Oct;48(1):1–9. doi: 10.1128/jvi.48.1.1-9.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Will H., Cattaneo R., Koch H. G., Darai G., Schaller H., Schellekens H., van Eerd P. M., Deinhardt F. Cloned HBV DNA causes hepatitis in chimpanzees. Nature. 1982 Oct 21;299(5885):740–742. doi: 10.1038/299740a0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES