Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1984 May;50(2):335–342. doi: 10.1128/jvi.50.2.335-342.1984

Expression of hepatitis B antigens with a simian virus 40 vector.

H Will, R Cattaneo, E Pfaff, C Kuhn, M Roggendorf, H Schaller
PMCID: PMC255625  PMID: 6323734

Abstract

Recombinant DNA molecules consisting of the simian virus 40 (SV40) early region and different subgenomic hepatitis B virus DNA fragments were constructed in vitro and packaged in vivo into SV40 capsids by using a complementing SV40 helper virus. Upon infection with these virus stocks the three known hepatitis B-specific antigens were expressed under SV40 control. The surface antigen was released into the medium, and the core antigen and its derivative hepatitis B e antigen were only detected intracellularly. Size analysis of the core gene product(s) by immunoblotting revealed the presence of a single protein species identical with the 21-kilodalton core antigen isolated from human liver. The hepatitis B core antigen expressing construct did not contain a putative precore sequence, indicating that such a sequence is not needed for hepatitis B core antigen synthesis in animal cells. S1 analysis demonstrated the use of SV40 signals for initiation and polyadenylation of the core gene transcripts. In addition, a processing-polyadenylation signal was identified within the core gene.

Full text

PDF
335

Images in this article

338Image
on p.338
339Image
on p.339

Selected References

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

  1. Cattaneo R., Will H., Darai G., Pfaff E., Schaller H. Detection of an element of the SV40 late promoter in vectors used for expression studies in COS cells. EMBO J. 1983;2(4):511–514. doi: 10.1002/j.1460-2075.1983.tb01455.x. [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. Christman J. K., Gerber M., Price P. M., Flordellis C., Edelman J., Acs G. Amplification of expression of hepatitis B surface antigen in 3T3 cells cotransfected with a dominant-acting gene and cloned viral DNA. Proc Natl Acad Sci U S A. 1982 Mar;79(6):1815–1819. doi: 10.1073/pnas.79.6.1815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen B. J., Richmond J. E. Electron microscopy of hepatitis B core antigen synthesized in E. coli. Nature. 1982 Apr 15;296(5858):677–679. doi: 10.1038/296677a0. [DOI] [PubMed] [Google Scholar]
  6. Colbère-Garapin F., Horodniceanu F., Kourilsky P., Garapin A. C. Late transient expression of human hepatitis B virus genes in monkey cells. EMBO J. 1983;2(1):21–25. doi: 10.1002/j.1460-2075.1983.tb01374.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Crowley C. W., Liu C. C., Levinson A. D. Plasmid-directed synthesis of hepatitis B surface antigen in monkey cells. Mol Cell Biol. 1983 Jan;3(1):44–55. doi: 10.1128/mcb.3.1.44. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cummings I. W., Browne J. K., Salser W. A., Tyler G. V., Snyder R. L., Smolec J. M., Summers J. Isolation, characterization, and comparison of recombinant DNAs derived from genomes of human hepatitis B virus and woodchuck hepatitis virus. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1842–1846. doi: 10.1073/pnas.77.4.1842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dejean A., Brechot C., Tiollais P., Wain-Hobson S. Characterization of integrated hepatitis B viral DNA cloned from a human hepatoma and the hepatoma-derived cell line PLC/PRF/5. Proc Natl Acad Sci U S A. 1983 May;80(9):2505–2509. doi: 10.1073/pnas.80.9.2505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dubois M. F., Pourcel C., Rousset S., Chany C., Tiollais P. Excretion of hepatitis B surface antigen particles from mouse cells transformed with cloned viral DNA. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4549–4553. doi: 10.1073/pnas.77.8.4549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Edman J. C., Hallewell R. A., Valenzuela P., Goodman H. M., Rutter W. J. Synthesis of hepatitis B surface and core antigens in E. coli. Nature. 1981 Jun 11;291(5815):503–506. doi: 10.1038/291503a0. [DOI] [PubMed] [Google Scholar]
  12. Feinman S. V., Overby L. R., Berris B., Chau K., Schable C. A., Maynard J. E. The significance of IgM antibodies to hepatitis B core antigen in hepatitis B carriers and hepatitis B-associated chronic liver disease. Hepatology. 1982 Nov-Dec;2(6):795–799. doi: 10.1002/hep.1840020609. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Ghosh P. K., Reddy V. B., Swinscoe J., Lebowitz P., Weissman S. M. Heterogeneity and 5'-terminal structures of the late RNAs of simian virus 40. J Mol Biol. 1978 Dec 25;126(4):813–846. doi: 10.1016/0022-2836(78)90022-0. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Gruss P., Khoury G. Expression of simian virus 40-rat preproinsulin recombinants in monkey kidney cells: use of preproinsulin RNA processing signals. Proc Natl Acad Sci U S A. 1981 Jan;78(1):133–137. doi: 10.1073/pnas.78.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Herrmann R., Neugebauer K., Pirkl E., Zentgraf H., Schaller H. Conversion of bacteriophage fd into an efficient single-stranded DNA vector system. Mol Gen Genet. 1980 Jan;177(2):231–242. doi: 10.1007/BF00267434. [DOI] [PubMed] [Google Scholar]
  19. Hirschman S. Z., Price P., Garfinkel E., Christman J., Acs G. Expression of cloned hepatitis B virus DNA in human cell cultures. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5507–5511. doi: 10.1073/pnas.77.9.5507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Koshy R., Koch S., von Loringhoven A. F., Kahmann R., Murray K., Hofschneider P. H. Integration of hepatitis B virus DNA: evidence for integration in the single-stranded gap. Cell. 1983 Aug;34(1):215–223. doi: 10.1016/0092-8674(83)90152-6. [DOI] [PubMed] [Google Scholar]
  21. Kozak M. Possible role of flanking nucleotides in recognition of the AUG initiator codon by eukaryotic ribosomes. Nucleic Acids Res. 1981 Oct 24;9(20):5233–5252. doi: 10.1093/nar/9.20.5233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  23. Liu C. C., Yansura D., Levinson A. D. Direct expression of hepatitis B surface antigen in monkey cells from an SV40 vector. DNA. 1982;1(3):213–221. doi: 10.1089/dna.1.1982.1.213. [DOI] [PubMed] [Google Scholar]
  24. MacKay P., Lees J., Murray K. The conversion of hepatitis B core antigen synthesized in E coli into e antigen. J Med Virol. 1981;8(4):237–243. doi: 10.1002/jmv.1890080404. [DOI] [PubMed] [Google Scholar]
  25. MacNab G. M., Alexander J. J., Lecatsas G., Bey E. M., Urbanowicz J. M. Hepatitis B surface antigen produced by a human hepatoma cell line. Br J Cancer. 1976 Nov;34(5):509–515. doi: 10.1038/bjc.1976.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  27. Miller R. H., Robinson W. S. Integrated hepatitis B virus DNA sequences specifying the major viral core polypeptide are methylated in PLC/PRF/5 cells. Proc Natl Acad Sci U S A. 1983 May;80(9):2534–2538. doi: 10.1073/pnas.80.9.2534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Moriarty A. M., Hoyer B. H., Shih J. W., Gerin J. L., Hamer D. H. Expression of the hepatitis B virus surface antigen gene in cell culture by using a simian virus 40 vector. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2606–2610. doi: 10.1073/pnas.78.4.2606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Piatak M., Subramanian K. N., Roy P., Weissman S. M. Late messenger RNA production by viable simian virus 40 mutants with deletions in the leader region. J Mol Biol. 1981 Dec 15;153(3):589–618. doi: 10.1016/0022-2836(81)90409-5. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  33. Roberts R. J., Breitmeyer J. B., Tabachnik N. F., Myers P. A. A second specific endonuclease from Haemophilus aegyptius. J Mol Biol. 1975 Jan 5;91(1):121–123. doi: 10.1016/0022-2836(75)90375-7. [DOI] [PubMed] [Google Scholar]
  34. Roggendorf M., Deinhardt F., Frösner G. G., Scheid R., Bayerl B., Zachoval R. Immunoglobulin M antibodies to hepatitis B core antigen: evaluation of enzyme immunoassay for diagnosis of hepatitis B virus infection. J Clin Microbiol. 1981 Apr;13(4):618–626. doi: 10.1128/jcm.13.4.618-626.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Siddiqui A. Expression of hepatitis B virus surface antigen gene in cultured cells by using recombinant plasmid vectors. Mol Cell Biol. 1983 Jan;3(1):143–146. doi: 10.1128/mcb.3.1.143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sompayrac L. M., Danna K. J. Efficient infection of monkey cells with DNA of simian virus 40. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7575–7578. doi: 10.1073/pnas.78.12.7575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  38. Stenlund A., Lamy D., Moreno-Lopez J., Ahola H., Pettersson U., Tiollais P. Secretion of the hepatitis B virus surface antigen from mouse cells using an extra-chromosomal eucaryotic vector. EMBO J. 1983;2(5):669–673. doi: 10.1002/j.1460-2075.1983.tb01482.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Stratowa C., Doehmer J., Wang Y., Hofschneider P. H. Recombinant retroviral DNA yielding high expression of hepatitis B surface antigen. EMBO J. 1982;1(12):1573–1578. doi: 10.1002/j.1460-2075.1982.tb01357.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Takahashi K., Akahane Y., Gotanda T., Mishiro T., Imai M., Miyakawa Y., Mayumi M. Demonstration of hepatitis B e antigen in the core of Dane particles. J Immunol. 1979 Jan;122(1):275–279. [PubMed] [Google Scholar]
  41. Takahashi K., Imai M., Gotanda T., Sano T., Oinuma A., Mishiro S., Miyakawa Y., Mayumi M. Hepatitis B e antigen polypeptides isolated from sera of individuals infected with hepatitis B virus: comparison with HBeAg polypeptide derived from Dane particles. J Gen Virol. 1980 Sep;50(1):49–57. doi: 10.1099/0022-1317-50-1-49. [DOI] [PubMed] [Google Scholar]
  42. Takahashi K., Imai M., Nomura M., Oinuma A., Machida A., Funatsu G., Miyakawa Y., Mayumi M. Demonstration of the immunogenicity of hepatitis B core antigen in a hepatitis B e antigen polypeptide (P19). J Gen Virol. 1981 Dec;57(Pt 2):325–330. doi: 10.1099/0022-1317-57-2-325. [DOI] [PubMed] [Google Scholar]
  43. 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]
  44. Varmus H. E. Form and function of retroviral proviruses. Science. 1982 May 21;216(4548):812–820. doi: 10.1126/science.6177038. [DOI] [PubMed] [Google Scholar]
  45. Wang Y., Schäfer-Ridder M., Stratowa C., Wong T. K., Hofschneider P. H. Expression of hepatitis B surface antigen in unselected cell culture transfected with recircularized HBV DNA. EMBO J. 1982;1(10):1213–1216. doi: 10.1002/j.1460-2075.1982.tb00015.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. 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]
  47. Will H., Kuhn C., Cattaneo R., Schaller H. Structure and function of the hepatitis B virus genome. Princess Takamatsu Symp. 1982;12:237–247. [PubMed] [Google Scholar]

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

RESOURCES