1887

Abstract

Hepatitis C virus (HCV) core protein forms the internal viral coat that encapsidates the genomic RNA and is enveloped in a host cell-derived lipid membrane. As the single capsid protein, core should be capable of multimerization but attempts to produce virus-like particles following expression of HCV structural proteins have not been successful. In this study, we have analysed the interaction capacity of full-length and truncated HCV core using the yeast two-hybrid system. Full-length core containing or lacking the translocation signal for the E1 glycoprotein did not interact with full-length or truncated core proteins. Truncation to the N-ter- minal 122 aa revealed an interaction domain which was mapped to the tryptophan-rich sequence from aa 82–102 and was termed the main homotypic interaction domain. The C-terminal hydrophobic fragment of core (aa 122–172) was incapable of interacting with itself but interacted with the main homotypic interaction domain in (the weak heterotypic interaction domain). Core proteins truncated at their N and C termini (aa 46-102) were trans-activating when fused to the DNA-binding domain of GAL4. Based on our results, we suggest that the C-terminal segment may interact in with the main homotypic interaction domain and thereby prevent multimerization. Core-core interaction was also observed for wtro-translated proteins bound to truncated immobilized core102. However, interaction was less specific in this system suggesting that protein interaction and possibly conformational alteration of core may be dependent on the experimental system.

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1997-06-01
2024-12-07
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References

  1. Abe K., Kurata T., Shikata T. 1989; Non-A, non-B hepatitis : visualization of virus-like particles from chimpanzee and human sera. Archives of Virology 104:351–355
    [Google Scholar]
  2. Bartel P. L., Chien C.-T., Sternglanz R., Fields S. 1993; Elimination of false positives that arise in using the two-hybrid system. Bio/Techniques 14:920–924
    [Google Scholar]
  3. Bartenschlager R., Ahlborn-Laake L., Mous J., Jacobsen H. 1993; Nonstructural protein 3 of the hepatitis C virus encodes a serine-type proteinase required for cleavage at the NS3/4 and NS4/5 junctions. Journal of Virology 67:3835–3844
    [Google Scholar]
  4. Birnbaum F., Nassal M. 1990; Hepatitis B virus nucleocapsid assembly: primary structure requirements in the core protein. Journal of Virology 64:3319–3330
    [Google Scholar]
  5. Bukh J., Purcell R. H., Miller R. 1994; Sequence analysis of the core gene of 14 hepatitis C virus genotypes. Proceedings of the National Academy of Sciences, USA 91:8239–8243
    [Google Scholar]
  6. Campbell S., Vogt V. M. 1995; Self-assembly in vitro of purified CA-NC proteins from Rous sarcoma virus and human immunodeficiency virus type 1. Journal of Virology 69:6487–6497
    [Google Scholar]
  7. Chang S. C., Yen J. H., Kang H. Y., Jang M. H., Chang M. F. 1994; Nuclear localization signals in the core protein of hepatitis C virus. Biochemical and Biophysical Research Communications 205:1284–1290
    [Google Scholar]
  8. Chien T.-C., Bartel P.L, Fields S. 1991; The two-hybrid system : a method to identify and clone genes for protein that interact with a protein of interest. Proceedings of the National Academy of Sciences, USA 88:9578–9582
    [Google Scholar]
  9. Choo Q.-L., Kuo G., Weiner A. J., Overby L. R., Bradley D. W., Houghton M. 1989; Isolation of cDNA clone derived from a blood- borne non-A, non-B viral hepatitis genome. Science 244:359–362
    [Google Scholar]
  10. Choo Q.-L., Richman K. H., Han J. H., Berger K., Lee C., Dong C., Gallegos C., Cott D., Edina-Selby A., Barr P. J., Weiner A. J., Bradley D. W., Kuo G., Houghton M. 1991; Genetic organization and diversity of the hepatitis C virus. Proceedings of the National Academy of Sciences, USA 88:2451–2455
    [Google Scholar]
  11. Durfee T., Becherer K., Chen P.-L., Yeh S.-H., Yang Y., Kilburn A., Lee W.-H., Elledge S. J. 1993; The retinoblastoma protein associated with the protein phosphatase type 1 catalytic subunit. Genes & Development 7:555–569
    [Google Scholar]
  12. Fields S., Song O. 1989; A novel genetic system to detect protein-protein interactions. Nature 340:245–247
    [Google Scholar]
  13. Francki R. I. B., Fauquet C. M., Knudson D. L., Brown F.editor 1991; Classification and Nomenclature of Viruses. Fifth Report of the International Committee on Taxonomy of Viruses. Archives of Virology Supplementum 2 p. 223
    [Google Scholar]
  14. Grakoui A., Wychowski C., Feinstone S. M., Rice C. M. 1993; Expression and identification of hepatitis C virus polyprotein cleavage products. Journal of Virology 67:1385–1395
    [Google Scholar]
  15. Harada S., Watanabe Y., Takeushi 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]
  16. Hijikata M., Kato N., Ootsuyama Y., Nakagawa M., Shimotohno K. 1991; Gene mapping of the putative structural region the hepatitis C virus genome by in vitro processing analysis. Proceedings of the National Academy of Sciences, USA 88:5547–5551
    [Google Scholar]
  17. Houghton M. 1996; Hepatitis C viruses. In Fields Virology, 3rd edn. pp. 1035–1058 Fields B. N., Knipe D. M., Howley P. M. Edited by Philadelphia: Lippincott-Raven;
    [Google Scholar]
  18. Kaito M., Watanabe S., Tsukiyama-Kohara K., Yamaguchi K., Kobayashi Y., Konishi M., Yokoi M., Ishida S., Suzuki S., Kohara M. 1994; Hepatitis C virus particle detected by immunoelectron microscopic study. Journal of General Virology 75:1755–1760
    [Google Scholar]
  19. 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 etiologic virus of human non-A, non-B hepatitis. Science 244:362–364
    [Google Scholar]
  20. Lanford R. E., Notvall L., Chavez D., White R., Frenzel G., Simonsen C., Kim J. 1993; Analysis of hepatitis C virus capsid, E1, and E2/NS1 proteins expressed in insect cells. Virology 197:225–235
    [Google Scholar]
  21. Legrain P., Dokhelar M.-C., Transy C. 1994; Detection of protein-protein interactions using different vectors in two-hybrid system. Nucleic Acids Research 22:3241–3242
    [Google Scholar]
  22. Li X., Jeffers L. J., Shao L., Reddy K. R., De Medina M., Scheffel J., Moore B., Schiff E. R. 1995; Identification of hepatitis C virus by immunelectron microscopy. Journal of Viral Hepatitis 2:227–234
    [Google Scholar]
  23. Lo S.-Y., Selby M., Tong M., Ou J.-H. 1994; Comparative studies of the core gene products of two different hepatitis C virus isolates: two alternative forms determined by a single amino acid substitution. Virology 199:124–131
    [Google Scholar]
  24. Lo S.-Y., Masiarz F., Hwang S. B., Lai M. M. C., Ou J.-H. 1995; Differential subcellular localization of hepatitis C virus core gene products. Virology 213:455–461
    [Google Scholar]
  25. Lu H.-H., Wimmer E. 1996; Poliovirus chimeras replicating under the translational control of genetic elements of hepatitis C virus reveal unusual properties of the internal ribosomal entry site of hepatitis C virus. Proceedings of the National Academy of Sciences, USA 93:1412–1417
    [Google Scholar]
  26. Luban J., Alin K. B., Bossolt K. L., Humaran T., Goff S. 1992; Genetic assay for multimerization of retroviral Gag polyproteins. Journal of Virology 66:5157–5160
    [Google Scholar]
  27. Matsumoto M., Hwang S. B., Jeng K.-S., Zhu N., Lai M. M. C. 1996; Homotypic interaction and multimerization of hepatitis C virus core protein. Virology 218:43–51
    [Google Scholar]
  28. Mizuno M., Yamada G., Tanaka T., Shimotohno K., Takatani M., Tsuji T. 1995; Virionlike structures in HeLa G cells transfected with the full-length sequence of the hepatitis C virus genome. Gastroenterology 109:1933–1940
    [Google Scholar]
  29. Moradpuor D., Englert C., Wakita T., Wands J. 1996; Characterization of cell lines allowing tightly regulated expression of hepatitis C virus core protein. Virology 222:51–63
    [Google Scholar]
  30. Mueller H. M., Pfaff E., Goeser T., Theilmann L. 1993; Genetic variability of German hepatitis C virus isolates. Journal of Medical Virology 40:291–306
    [Google Scholar]
  31. Ray R. B., Lagging L. M., Meyer K., Steele R., Ray R. 1995; Transcriptional regulation of cellular and viral promoters by the hepatitis C virus core protein. Virus Research 37:209–220
    [Google Scholar]
  32. Rice C. 1996; Flaviviridae : the viruses and their replication. In Fields Virology, 3rd edn. pp. 931–959 Fields B. N., Knipe D. M., Howley P. M. Edited by Philadelphia: Lippincott-Raven;
    [Google Scholar]
  33. Santolini E., Migliaccio G., La Monica N. 1994; Biosynthesis and biochemical properties of the hepatitis C virus core protein. Journal of Virology 68:3631–3641
    [Google Scholar]
  34. Shih C.-M., Lo S. J., Miyamura T., Chen S.-Y., Lee Y.-H. W. 1993; Suppression of hepatitis B virus expression and replication by hepatitis C virus core protein in HuH-7 cells. Journal of Virology 67:5823–5832
    [Google Scholar]
  35. Shih C.-M., Chen C.-M., Chen S.-Y., Lee Y.-H. W. 1995; Modulation of the trans-suppression activity of hepatitis C virus core protein by phosphorylation. Journal of Virology 69:1160–1171
    [Google Scholar]
  36. Shimizu Y. K., Feinstone S. M., Kohara M., Purcell R. H., Yoshikura H. 1996; Hepatitis C virus : detection of intracellular virus particles by electron microscopy. Hepatology 23:205–209
    [Google Scholar]
  37. Suzuki R., Matsuura Y., Suzuki T., Ando A., Chiba J., Harada S., Saito I., Miamura T. 1995; Nuclear localization of the truncated hepatitis C virus core protein with its hydrophobic C-terminus deleted. Journal of General Virology 76:53–61
    [Google Scholar]
  38. Takahashi K., Kishimoto S., Yoshizawa H., Okamoto H., Yoshikawa A., Mishiro S. 1992; p26 protein and 33nm particle associated with nucleocapsid of hepatitis C virus recovered from the circulation of infected hosts. Virology 191:431–434
    [Google Scholar]
  39. Takamizawa A., Mori C., Fuke I., Manabe S., Murakami S., Fujita J., Onushi 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]
  40. Takeuchi K., Kubo Y., Boonmar S., Watanabe Y., Kakayama T., Choo Q.-L., Kuo G., Houghton M., Saito I., Miyamura T. 1990; The putative nucleocapsid and envelope protein genes of the hepatitis C virus determined by comparison of two isolates derived from an experimentally infected chimpanzee and healthy human carriers. Journal of General Virology 71:3027–3033
    [Google Scholar]
  41. Tsukiyama-Kohara K., Iizuka N., Kohara M., Nomoto A. 1992; Internal ribosome entry site within hepatitis C virus RNA. Journal of Virology 66:1476–1483
    [Google Scholar]
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