Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1985 Jun;4(6):1507–1514. doi: 10.1002/j.1460-2075.1985.tb03810.x

Transcription of woodchuck hepatitis virus in the chronically infected liver.

T Möröy, J Etiemble, C Trépo, P Tiollais, M A Buendia
PMCID: PMC554375  PMID: 4029122

Abstract

The transcription of woodchuck hepatitis virus (WHV) genome was studied in the liver of chronically infected woodchucks by Northern blot, nuclease mapping and primer extension analysis. Two major transcripts, 2.1 and 3.7 kb in length, and several minor transcripts were found in samples which supported active WHV replication. The 2.1-kb RNA represents the major transcript of the S gene, encoding the viral surface antigen (WHsAg) as demonstrated by blot-hybridization experiments. Two transcription initiation sites were localized downstream of the second AUG of the pre-S region, 139 and 152 nucleotides upstream of the translation initiation codon of the S gene. The 3.7-kb transcript, present in an equal amount, is slightly larger than the WHV genome and could be involved in the expression of all viral proteins. The data derived from RNA mapping strongly suggest that this transcript is initiated approximately 70 nucleotides upstream of the C gene, encoding the viral core antigen (WHcAg), and represents the message for WHcAg. It might also serve in the viral replication cycle as a potential template for reverse transcription. All WHV-specific transcripts were found to be processed at a unique site, 20 nucleotides downstream of the polyadenylation signal situated within the core gene. A different set of WHV-specific mRNAs was observed in a woodchuck hepatocellular carcinoma when only integrated forms of WHV DNA could be detected. Two RNA species of 2.3 and 4.6 kb were characterized. The 3.7-kb RNA was absent, reinforcing the hypothesis that this transcript corresponds to the pre-genome.

Full text

PDF
1507

Images in this article

1508Fig. 1.
on p.1508
1509Fig. 2.
on p.1509
1510Fig. 3.
on p.1510
1511Fig. 4.
on p.1511
1512Fig. 5.
on p.1512

Selected References

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

  1. Auffray C., Rougeon F. Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA. Eur J Biochem. 1980 Jun;107(2):303–314. doi: 10.1111/j.1432-1033.1980.tb06030.x. [DOI] [PubMed] [Google Scholar]
  2. Berk A. J., Sharp P. A. Spliced early mRNAs of simian virus 40. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1274–1278. doi: 10.1073/pnas.75.3.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brechot C., Pourcel C., Louise A., Rain B., Tiollais P. Presence of integrated hepatitis B virus DNA sequences in cellular DNA of human hepatocellular carcinoma. Nature. 1980 Jul 31;286(5772):533–535. doi: 10.1038/286533a0. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Cattaneo R., Will H., Schaller H. Hepatitis B virus transcription in the infected liver. EMBO J. 1984 Sep;3(9):2191–2196. doi: 10.1002/j.1460-2075.1984.tb02113.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chakraborty P. R., Ruiz-Opazo N., Shouval D., Shafritz D. A. Identification of integrated hepatitis B virus DNA and expression of viral RNA in an HBsAg-producing human hepatocellular carcinoma cell line. Nature. 1980 Jul 31;286(5772):531–533. doi: 10.1038/286531a0. [DOI] [PubMed] [Google Scholar]
  7. Dejean A., Sonigo P., Wain-Hobson S., Tiollais P. Specific hepatitis B virus integration in hepatocellular carcinoma DNA through a viral 11-base-pair direct repeat. Proc Natl Acad Sci U S A. 1984 Sep;81(17):5350–5354. doi: 10.1073/pnas.81.17.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Edman J. C., Gray P., Valenzuela P., Rall L. B., Rutter W. J. Integration of hepatitis B virus sequences and their expression in a human hepatoma cell. Nature. 1980 Jul 31;286(5772):535–538. doi: 10.1038/286535a0. [DOI] [PubMed] [Google Scholar]
  9. Favaloro J., Treisman R., Kamen R. Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 1980;65(1):718–749. doi: 10.1016/s0076-6879(80)65070-8. [DOI] [PubMed] [Google Scholar]
  10. Feitelson M. A., Marion P. L., Robinson W. S. Antigenic and structural relationships of the surface antigens of hepatitis B virus, ground squirrel hepatitis virus, and woodchuck hepatitis virus. J Virol. 1981 Aug;39(2):447–454. doi: 10.1128/jvi.39.2.447-454.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Fowler M. J., Monjardino J., Tsiquaye K. N., Zuckerman A. J., Thomas H. C. The mechanism of replication of hepatitis B virus: evidence of asymmetric replication of the two DNA strands. J Med Virol. 1984;13(1):83–91. doi: 10.1002/jmv.1890130110. [DOI] [PubMed] [Google Scholar]
  13. Frommel D., Crevat D., Vitvitsky L., Pichoud C., Hantz O., Chevalier M., Grimaud J. A., Lindberg J., Trépo C. G. Immunopathologic aspects of woodchuck hepatitis. Am J Pathol. 1984 Apr;115(1):125–134. [PMC free article] [PubMed] [Google Scholar]
  14. Gough N. M. Core and E antigen synthesis in rodent cells transformed with hepatitis B virus DNA is associated with greater than genome length viral messenger RNAs. J Mol Biol. 1983 Apr 25;165(4):683–699. doi: 10.1016/s0022-2836(83)80274-5. [DOI] [PubMed] [Google Scholar]
  15. Gough N. M., Murray K. Expression of the hepatitis B virus surface, core and E antigen genes by stable rat and mouse cell lines. J Mol Biol. 1982 Nov 25;162(1):43–67. doi: 10.1016/0022-2836(82)90161-9. [DOI] [PubMed] [Google Scholar]
  16. Laub O., Rall L. B., Truett M., Shaul Y., Standring D. N., Valenzuela P., Rutter W. J. Synthesis of hepatitis B surface antigen in mammalian cells: expression of the entire gene and the coding region. J Virol. 1983 Oct;48(1):271–280. doi: 10.1128/jvi.48.1.271-280.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Machida A., Kishimoto S., Ohnuma H., Miyamoto H., Baba K., Oda K., Nakamura T., Miyakawa Y., Mayumi M. A hepatitis B surface antigen polypeptide (P31) with the receptor for polymerized human as well as chimpanzee albumins. Gastroenterology. 1983 Aug;85(2):268–274. [PubMed] [Google Scholar]
  18. Mandart E., Kay A., Galibert F. Nucleotide sequence of a cloned duck hepatitis B virus genome: comparison with woodchuck and human hepatitis B virus sequences. J Virol. 1984 Mar;49(3):782–792. doi: 10.1128/jvi.49.3.782-792.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. Michel M. L., Pontisso P., Sobczak E., Malpièce Y., Streeck R. E., Tiollais P. Synthesis in animal cells of hepatitis B surface antigen particles carrying a receptor for polymerized human serum albumin. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7708–7712. doi: 10.1073/pnas.81.24.7708. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Miller R. H., Tran C. T., Robinson W. S. Hepatitis B virus particles of plasma and liver contain viral DNA-RNA hybrid molecules. Virology. 1984 Nov;139(1):53–63. doi: 10.1016/0042-6822(84)90329-5. [DOI] [PubMed] [Google Scholar]
  23. Ogston C. W., Jonak G. J., Rogler C. E., Astrin S. M., Summers J. Cloning and structural analysis of integrated woodchuck hepatitis virus sequences from hepatocellular carcinomas of woodchucks. Cell. 1982 Jun;29(2):385–394. doi: 10.1016/0092-8674(82)90155-6. [DOI] [PubMed] [Google Scholar]
  24. Ponzetto A., Cote P. J., Ford E. C., Purcell R. H., Gerin J. L. Core antigen and antibody in woodchucks after infection with woodchuck hepatitis virus. J Virol. 1984 Oct;52(1):70–76. doi: 10.1128/jvi.52.1.70-76.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pourcel C., Louise A., Gervais M., Chenciner N., Dubois M. F., Tiollais P. Transcription of the hepatitis B surface antigen gene in mouse cells transformed with cloned viral DNA. J Virol. 1982 Apr;42(1):100–105. doi: 10.1128/jvi.42.1.100-105.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rall L. B., Standring D. N., Laub O., Rutter W. J. Transcription of hepatitis B virus by RNA polymerase II. Mol Cell Biol. 1983 Oct;3(10):1766–1773. doi: 10.1128/mcb.3.10.1766. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rogler C. E., Summers J. Novel forms of woodchuck hepatitis virus DNA isolated from chronically infected woodchuck liver nuclei. J Virol. 1982 Dec;44(3):852–863. doi: 10.1128/jvi.44.3.852-863.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. Simonsen C. C., Levinson A. D. Analysis of processing and polyadenylation signals of the hepatitis B virus surface antigen gene by using simian virus 40-hepatitis B virus chimeric plasmids. Mol Cell Biol. 1983 Dec;3(12):2250–2258. doi: 10.1128/mcb.3.12.2250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. 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]
  32. 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]
  33. 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]
  34. 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]
  35. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Weaver R. F., Weissmann C. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates. Nucleic Acids Res. 1979 Nov 10;7(5):1175–1193. doi: 10.1093/nar/7.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Will H., Cattaneo R., Pfaff E., Kuhn C., Roggendorf M., Schaller H. Expression of hepatitis B antigens with a simian virus 40 vector. J Virol. 1984 May;50(2):335–342. doi: 10.1128/jvi.50.2.335-342.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES