1887

Abstract

We have expressed the full-length coding region and selected domains of the hepatitis C virus (HCV) cDNA in mammalian cells by transfection. Using HCV antibody-positive human sera and monospecific antibodies the proteins encoded by the putative structural and non-structural regions of the open reading frame of HCV were identified as core (p22), E1 (gp32–35), E2 (gp68–72), NS2 (p23), NS3 (p72), NS4a and b (p10 and p27) and NS5a and b (p56 and p70). We have also defined the subcellular localizations of the HCV proteins using indirect immunofluorescence assays.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-74-6-1103
1993-06-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/74/6/JV0740061103.html?itemId=/content/journal/jgv/10.1099/0022-1317-74-6-1103&mimeType=html&fmt=ahah

References

  1. Chambers T. J., Hahn C. S., Galler R., Rice C. M. 1990; Flavivirus genome organization, expression, and replication. Annual Review of Microbiology 44:649–688
    [Google Scholar]
  2. Choo Q.-L., Kuo G., Weiner A. J., Overby L. J., Bradley D. W., Houghton M. 1989; Isolation of a cDNA clone derived from blood-borne nonA, nonB viral hepatitis genome. Science 244:359–362
    [Google Scholar]
  3. Choo Q.-L., Weiner A. J., Overby L. R., Kuo G., Houghton M., Bradley D. W. 1990; Hepatitis C virus: the major causative agent of viral non-A, non-B hepatitis. British Medical Bulletin 46:423–441
    [Google Scholar]
  4. Choo Q.-L., Han J., Weiner A. J., Overby L. R., Bradley D. W., Kuo G., Houghton M. 1991a; Hepatitis C virus is a distant relative of the flaviviruses and pestiviruses. In Viral Hepatitis C, D and E pp. 47–52 Edited by Shikata T., Purcell R. H., Uchida T. Amsterdam: Elsevier;
    [Google Scholar]
  5. Choo Q.-L., Richman K. H., Han J. H., Berger K., Lee C., Dong C., Gallegos C., Coit D., Medina-Selby A., Barr P. J., Weiner A. J., Bradley D. W., Kuo G., Houghton M. 1991b; Genetic organization and diversity of the hepatitis C virus. Proceedings of the National Academy of Sciences, U.S.A. 88:2451–2455
    [Google Scholar]
  6. Collett M. S., Larson R., Gold C., Strick D., Anderson D. K., Purchio A. F. 1988a; Molecular cloning and nucleotide sequence of the pestivirus bovine viral diarrhea virus. Virology 165:191–199
    [Google Scholar]
  7. Collett M. S., Larson R., Belzer S. K., Retzel E. 1988b; Proteins encoded by bovine viral diarrhea virus: the genomic organization of a pestivirus. Virology 165:200–208
    [Google Scholar]
  8. Donahue J. G., Munoz A., Ness P. M., Brown D. E. Jr, Yawn D. H., McAllister H. A., Reitz B. A., Nelson K. E. 1992; The declining risk of post-transfusion hepatitis C virus infection. New England Journal of Medicine 327:371–373
    [Google Scholar]
  9. Elroy-Stein O., Moss B. 1990; Cytoplasmic expression system based on constitutive synthesis of bacteriophage T7 RNA polymerase in mammalian cells. Proceedings of the National Academy of Sciences, U.S.A. 87:6743–6747
    [Google Scholar]
  10. Han J. H., Shyamala V., Richman K., Brauer M. J., Irvine B., Urdea M. S., Tekamp-Olson P., Kuo G., Choo Q.-L., Houghton M. 1991; Characterization of the terminal regions of hepatitis C viral RNA: identification of conserved sequences in the 5′ untranslated region and poly(A) tails at the 3′ end. Proceedings of the National Academy of Sciences, U.S.A. 88:1711–1715
    [Google Scholar]
  11. Harada S., Watanabe Y., Takeuchi K., Suzuki T., Katayama T., Takebe Y., Saito I., Miyamura T. 1991; Expression of processed core protein of hepatitis C virus in mammalian cells. Journal of Virology 65:3015–3021
    [Google Scholar]
  12. Hijikata M., Kato N., Ootsuyama Y., Nakagawa M., Shimotohno K. 1991; Gene mapping of the putative structural region of the hepatitis C virus genome by in vitro processing analysis. Proceedings of the National Academy of Sciences, U.S.A. 88:5547–5551
    [Google Scholar]
  13. Houghton M. 1991; Molecular virology of HCV. In Proceedings of the Second International Symposium on HCV pp. 2–3 New Jersey: Advanced Therapeutics Communications;
    [Google Scholar]
  14. Houghton M., Weiner A., Han J., Kuo G., Choo Q.-L. 1992; Heterogeneity of the HCV genome: importance for control of the disease. In Hepatitis C Virus: Scientific and Clinical Status pp. 8–9 Edited by Deinhardt F., Bradley D. W., Houghton M. New Jersey: Advanced Therapeutic Communications;
    [Google Scholar]
  15. Kamer G., Argos P. 1984; Primary structural composition of RNA-dependent polymerases from plant, animal and bacterial viruses. Nucleic Acids Research 12:7269–7282
    [Google Scholar]
  16. Kato N., Hijikata M., Ootsuyama Y., Nakagawa M., Ohkoshi S., Sugimura T., Shimotohno K. 1990; Molecular cloning of the human hepatitis C virus genome from Japanese patients with non-A, non-B hepatitis. Proceedings of the National Academy of Sciences, U.S.A. 87:9524–9528
    [Google Scholar]
  17. Kitamura N., Semler B., Rothberg P. G., Larsen G. R., Adler C. J., Dorner A. J., Emine E. A., Hanecak R., Lee J. L., Werf S., Anderson C. W., Wimmbr E. 1981; Primary structure, gene organization and polypeptide expression of poliovirus RNA. Nature, London 291:547–553
    [Google Scholar]
  18. Krawczynski K., Beach M. J., Bradley D. W., Kuo G., di Bisceglie A. M., Houghton M., Reyes G. R., Kim J. P., Choo Q.-L., Alter M. J. 1992; Hepatitis C virus antigen in hepatocytes: immunomorphogenic detection and identification. Gastroenterology 103:622–629
    [Google Scholar]
  19. Krzysztof K., Beach M. J., Bradley D. W., Kuo G., di Bisceglie A. M., Houghton M., Reyes G. R., Kim J. P., Choo Q.-L., Alter M. J. 1992; Hepatitis C virus antigen in hepatocytes: immunomorphologic detection and identification. Gastroenterology 103:622–629
    [Google Scholar]
  20. Kumar U., Cheng D., Thomas H., Monjardino J. 1992; Cloning and sequencing of the structural region and expression of putative core gene of hepatitis C virus from a British case of chronic sporadic hepatitis. Journal of General Virology 73:1521–1525
    [Google Scholar]
  21. Kuo G., Choo Q.-L., Alter H. J., Gitnick G. L., Redeker A. G., Purcell R. H., Miyamura T., Dienstag J. L., Alter M. J., Stevens C. E., Tegtmeier G. E., Bonino F., Colombo M., Lee W.-S., Kuo C., Berger K., Shuster J. R., Overby L. R., Bradley D. W., Houghton M. 1989; An assay for circulating antibodies to a major etiologie virus of human non-A, non-B hepatitis. Science 244:362–364
    [Google Scholar]
  22. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  23. Matsuura Y., Harada S., Suzuki R., Watanabe Y., Inoue Y., Saito I., Miyamura T. 1992; Expression of processed envelope protein of hepatitis C virus in mammalian and insect cells. Journal of Virology 66:1425–1431
    [Google Scholar]
  24. Okamoto H., Okada S., Sugiyama Y., Kurai K., Iizuka H., Machida A., Miyakawa Y., Mayumi M. 1991; Nucleotide sequence of the genomic RNA of hepatitis C virus isolated from a human carrier: comparison with reported isolates for conserved and divergent regions. Journal of General Virology 72:2697–2704
    [Google Scholar]
  25. Rice C. M., Strauss E. G., Strauss J. H. 1986; Structure of the flavivirus genome. In The Togaviridae and Flaviviridae pp. 279–326 Edited by Schlesinger S., Schlesinger M. J. New York: Plenum Press;
    [Google Scholar]
  26. Spaete R. R., Alexander D., Rugroden M. E., Choo Q.-L., Berger K., Crawford K., Kuo C., Leng S., Lee C., Ralston R., Thudium K., Tung J. W., Kuo G., Houghton M. 1992; Characterization of the hepatitis C virus E2/NS1 gene product expressed in mammalian cells. Virology 188:819–830
    [Google Scholar]
  27. Takamizawa A., Mori C., Fuke I., Manabe S., Murakami S., Fujita J., Onishi E., Andoh T., Yoshida I., Okayama H. 1991; Structure and organization of the hepatitis C virus genome isolated from human carriers. Journal of Virology 65:1105–1113
    [Google Scholar]
  28. Takeuchi K., Kubo Y., Boonmar S., Watanabe Y., Katayama T., Choo Q.-L., Kuo G., Houghton M., Saito I., Miyamura T. 1990; The putative nucleocapsid and envelope protein genes of hepatitis C virus determined by comparison of the nucleocapsid sequences of two isolates derived from an experimentally infected chimpanzee and healthy human carriers. Journal of General Virology 71:3027–3033
    [Google Scholar]
  29. Weiner A. J., Brauer M. J., Rosenblatt J., Richman K. H., Tung J., Crawford K., Bonino F., Saracco G., Choo Q.-L., Houghton M., Han J. H. 1991; Variable and hypervariable domains are found in the regions of HCV corresponding to the flavivirus envelope and NS1 proteins and the pestivirus envelope glycoproteins. Virology 180:842–848
    [Google Scholar]
  30. Yoo B. J., Spaete R., Gebale A. P., Selby M., Houghton M., Han J. H. 1992; 5′ end-dependent translation initiation of hepatitis C viral RNA and the presence of putative positive and negative translational control elements within the 5′ untranslated region. Virology 191:889–899
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-74-6-1103
Loading
/content/journal/jgv/10.1099/0022-1317-74-6-1103
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error