1887

Abstract

SUMMARY

The core protein of hepatitis C virus (HCV) is considered to be cleaved from the N terminus of the large precursor polyprotein by cellular signalase. The HCV cDNA encoding the core protein was expressed (i) in monkey COS cells by a plasmid expression vector driven by the SRα promoter, and (ii) in insect cells by a recombinant baculovirus. The expressed product had an of 22000 and was located in the cytoplasm. When the C-terminal hydrophobic domains were deleted, however, the truncated core proteins were translocated into the nucleus. The truncated core proteins were located in the nucleus even when they were expressed as a fusion protein with . -galactosidase, which is essentially localized in the cytoplasm. Plasmids containing HCV cDNAs with a deletion in one of the regions encoding clusters of basic amino acids were expressed in COS cells and the localization of the core protein was examined. The residues PRRGPR were suggested to play an important role in nuclear localization. HCV is an RNA virus and its life cycle was originally considered to be confined to the cytoplasm; the present study, however, suggests that the HCV core protein can translocate into the nucleus under certain circumstances.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-76-1-53
1995-01-01
2022-01-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/76/1/JV0760010053.html?itemId=/content/journal/jgv/10.1099/0022-1317-76-1-53&mimeType=html&fmt=ahah

References

  1. Bruix J., Barrera J. M., Calvet X., Ercilla G., Costa J., Sanchez-Tapias J. M., Ventura M., Vall M., Bruguera M., Bru C., Castillo R., Rodes J. 1989; Prevalence of antibodies to hepatitis C virus in Spanish patients with hepatocellular carcinoma and hepatic cirrhosis. Lancet ii:1004–1006
    [Google Scholar]
  2. Bulich R., Aaskov J. G. 1992; Nuclear localization of dengue 2 virus core protein detected with monoclonal antibodies. Journal of General Virology 73:2999–3003
    [Google Scholar]
  3. Chiba J., Ohba H., Matsuura Y., Watanabe Y., Katayama T., Kikuchi S., Saito I., Miyamura T. 1991; Serodiagnosis of hepatitis C virus (HCV) infection with an HCV core protein molecularly expressed by a recombinant baculovirus. Proceedings of the National Academy of SciencesUSA 88:4641–4645
    [Google Scholar]
  4. Choo Q. -L., Kuo G., Weiner A. J., Overby L. R., Bradley D. W., Houghton M. 1989; Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science 244:359–362
    [Google Scholar]
  5. Choo Q. -L., Richman K., Han J., 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. 1991; Genetic organization and diversity of the hepatitis C virus. Proceedings of the National Academy of SciencesUSA 88:2451–2455
    [Google Scholar]
  6. Chou W. -H., Yoneyama T., Takeuchi K., Saito I., Miyamura T. 1991; Discrimination of hepatitis C virus in liver tissues from different patients with hepatocellular carcinomas by direct nucleotide sequencing of amplified cDNA of the viral genome. Journal of Clinical Microbiology 29:2860–2864
    [Google Scholar]
  7. Colombo M., Kuo G., Choo Q. -L., Donato M. F., Ninno E. D., Tommasini M. A., Dioguardi N., Houghton M. 1989; Prevalence of antibodies to hepatitis C virus in Italian patients with hepatocellular carcinoma. Lancet ii:1006–1008
    [Google Scholar]
  8. Feldherr C. M., Kallenbach E., Schultz N. 1984; Movement of a karyophilic protein through the nuclear pores of oocytes. Journal of Cell Biology 99:2216–2222
    [Google Scholar]
  9. Grakoui A., McCourt D. W., Wychowski C., Feinstone S. M., Rice C. M. 1993a; Characterization of the hepatitis C virus- encoded serine proteinase: determination of proteinase-dependent polyprotein cleavage sites. Journal of Virology 67:2832–2843
    [Google Scholar]
  10. Grakoui A., Wychowski C., Lin C., Feinstone S. M., Rice C. M. 1993b; Expression and identification of hepatitis C virus polyprotein cleavage products. Journal of Virology 67:1385–1395
    [Google Scholar]
  11. Han J. H., Shyamala K. H., Richman K. H., 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 SciencesUSA 88:171–1715
    [Google Scholar]
  12. Harada S., Takeuchi K., Suzuki T., Katayama T., Watanabe Y., 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]
  13. 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 SciencesUSA 88:5547–5551
    [Google Scholar]
  14. Hiramatsu N., Hayashi N., Haruna Y., Kasahara A., Fusamoto H., Mori C., Fuke I., Okayama H., Kamada T. 1992; Immunohistochemical detection of hepatitis C virus-infected hepato- cytes in chronic liver disease with monoclonal antibodies to core, envelope and NS3 regions of the hepatitis C virus genome. Hepatology 16:306–311
    [Google Scholar]
  15. Hosein B., Fang C. T., Popovsky M. A., Ye J., Zhang M., Wang C. Y. 1991; Improved serodiagnosis of hepatitis C virus infection with synthetic peptide antigen from capsid protein. Proceedings of the National Academy of SciencesUSA 88:3647–3651
    [Google Scholar]
  16. Houghton M., Weiner A., Han J., Kuo G., Choo Q. -L. 1991; Molecular biology of the hepatitis C viruses: implications for diagnosis, development and control of viral disease. Hepatology 14:381–388
    [Google Scholar]
  17. Kalderon D., Roberts B. L., Richardson W. D., Smith A. E. 1984; A short amino acid sequence able to specify nuclear location. Cell 39:499–509
    [Google Scholar]
  18. Katayama T., Matsuda T., Kikuchi S., Harada S., Matsuura Y., Chiba J., Saito I., Miyamura T. 1992; Improved serodiagnosis of non-A, non-B hepatitis by an assay detecting antibody to hepatitis C virus core antigen. Hepatology 15:391–394
    [Google Scholar]
  19. 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 SciencesUSA 87:9524–9528
    [Google Scholar]
  20. Kim D. -W., Suzuki R., Ando A., Harada S., Matsuura Y., Harada T., Chiba J., Saito I., Miyamura T. 1991; Domains influencing the intracellular localization of hepatitis C virus core protein. The 1991 Meeting on Molecular Biology of Hepatitis B Viruses Paris:213
    [Google Scholar]
  21. Kohara M., Tsukiyama-Kohara K., Maki N., Asano K., Yamaguchi K., Miki K., Tanaka S., Hattori N., Matsuura Y., Saito I., Miyamura T., Nomoto A. 1992; Expression and characterization of glycoprotein gp35 of hepatitis C virus using recombinant vaccinia virus. Journal of General Virology 73:2313–2318
    [Google Scholar]
  22. Kubo Y., Takeuchi K., Boonmar S., Katayama T., Choo Q. -L., Kuo G., Weiner A. W., Bradley D. W., Houghton M., Saito I., Miyamura T. 1989; A cDNA fragment of hepatitis C virus isolated from an implicated donor of post-transfusion non-A, non-B hepatitis in Japan. Nucleic Acids Research 17:10367–10372
    [Google Scholar]
  23. 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 F. 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 244:362364
    [Google Scholar]
  24. Kyte J., Doolittle R. F. 1982; A simple method for displaying the hydropathic character of a protein. Journal of Molecular Biology 157:105–132
    [Google Scholar]
  25. Lanford R. E., Notvall L., Chavez D., White R., Frenzel G., Simonsen C., Kim J. 1993; Analysis of hepatitis C virus capsid, El, and E2/NS1 proteins expressed in insect cells. Virology 197:225–235
    [Google Scholar]
  26. 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 124:124–131
    [Google Scholar]
  27. Maekawa T., Matsuda S., Fujisawa J., Yoshida M., Ishii S. 1991; Cyclic AMP response element-binding protein, CRE-BP1, mediates the ElA-induced but not the Tax-induced trans-activation. Oncogene 6:627–632
    [Google Scholar]
  28. Manabe S., Fuke I., Tanishita O., Kaji C., Gomi Y., Yoshida S., Mori C., Takamizawa A., Yoshida I., Okayama H. 1994; Production of nonstructural proteins of hepatitis C virus requires a putative viral protease encoded by NS3. Virology 198:636–644
    [Google Scholar]
  29. Martynoff G. -D., Venneman A., Wyseur A., Maertens G. 1993; Characterization of recombinant vaccinia viruses expressing core/El proteins of hepatitis C virus. International Symposium on Viral Hepatitis and Liver Disease Tokyo: 292173
    [Google Scholar]
  30. Matsuura Y., Possee R. D., Overton H. A., Bishop D. H. L. 1987; Baculovirus expression vectors: the requirements for high level expression of proteins, including glycoproteins. Journal of General Virology 68:1233–1250
    [Google Scholar]
  31. 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]
  32. Matsuura Y., Suzuki R., Watanabe Y., Suzuki T., Miyamura T. 1993; Expression of E1 and E2/NS1 proteins of HCV in mammalian and insect cells. In Vaccine ’93: Modern Approaches to New Vaccines including Prevention of AIDS289–294 Brown F., Chanock R. M., Ginsberg H. S., Lerner R. A. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  33. Miller R. H., Purcell R. H. 1990; Hepatitis C virus shares amino acid sequence similarity with pestiviruses and flaviviruses as well as members of two plant virus supergroups. Proceedings of the National Academy of SciencsUSA 87:2057–2061
    [Google Scholar]
  34. Moreland R. B., Nam H. C., Hereford L. M., Fried H. M. 1985; Identification of a nuclear localization signal of a yeast ribosomal protein. Proceedings of the National Academy of SciencesUSA 82:6561–6565
    [Google Scholar]
  35. Mori S., Kato N., Yagyu A., Tanaka T., Ikeda Y., Petchclai B., Chiewsilp P., Kurimura T., Shimotohno K. 1992 A new type of hepatitis C virus in patients in Thailand. Biochemical and Biophysical Research Communications 183:334–342
    [Google Scholar]
  36. Nishioka K., Watanabe J., Furuta S., Tanaka E., Iino S., Suzuki H., Tsuji T., Yano M., Kuo G., Choo Q. -L., Houghton M., Oda T. 1991; A high prevalence of antibody to the hepatitis- C virus in patients with hepatocellular carcinoma in Japan. Cancer 67:429–433
    [Google Scholar]
  37. Nowak T., Farber P. M., Wengler G., Wengler G. 1989; Analyses of the terminal sequences of West Nile virus structural proteins and of thein vitro translation of these proteins allow the proposal of a complete scheme. Virology 169:365–376
    [Google Scholar]
  38. Okamoto H., Kurai K., Okada S., Yamamoto K., Lizuka H., Tanaka T., Fukuda S., Tsuda F., Mishiro S. 1992; Full- length sequence of a hepatitis C virus genome having poor homology to reported isolates: comparative study of four distinct genotypes. Virology 188:331–341
    [Google Scholar]
  39. Paine P. L., Moore L. C., Horowitz S. B. 1975; Nuclear envelope permeability. Nature 254:109–114
    [Google Scholar]
  40. Perry L. J., Rixon F. J., Everett R. D., Frame M. C., McGeoch D. J. 1986; Characterization of the IE110 gene of herpes simplex virus type 1. Journal of General Virology 67:2365–2380
    [Google Scholar]
  41. Ravaggi A., Natoli G., Primi D., Albertini A., Levrero M., Caniari E. 1994; Intranuclear localization of full-length and truncated hepatitis C virus core protein expressed in mammalian cells. Journal of Hepatology 20:833–836
    [Google Scholar]
  42. Rihs H. -P., Jans D. A., Fan H., Peters R. 1991; The rate of nuclear cytoplasmid protein transport is determined by the casein kinase II site flanking the nuclear localization sequence of the SV40 T-antigen. EMBO Journal 10:633–639
    [Google Scholar]
  43. Rogel-Gaillard C., Breitburd F., Orth G. 1992; Human papillomavirus type 1 E4 proteins differing by their N-terminal ends have distinct cellular localizations when transiently expressedin vitro. Journal of Virology 66:816–823
    [Google Scholar]
  44. Saito I., Miyamura T., Ohbayashi A., Harada H., Katayama T., Kikuchi S., Watanabe Y., Koi S., Onji M., Ohta Y., Choo Q. -L., Houghton M., Kuo G. 1990; Hepatitis C virus infection is associated with the development of hepatocellular carcinoma. Proceedings of the National Academy of SciencesUSA 87:6547–6549
    [Google Scholar]
  45. Sarashina T., Sakurai T., Watanabe Y., Kashima K., Suzuki T., Chiba J., Kita Y., Horiuchi T., Saito I., Miyamura T. 1993; Nucleotide sequence of the hepatitis C virus genome from a patient negative for anti-HCV by the first generation antibody assay. Nucleic Acids Research 21:1037
    [Google Scholar]
  46. Selby M. J., Choo Q.-L., Berger G., Glazer E., Eckart M., Lee C., Chien D., Kuo C., Houghton M. 1993; Expression, identification and subcellular localization of the proteins encoded by the hepatitis C viral genome. Journal of General Virology 74:1103–1113
    [Google Scholar]
  47. 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]
  48. Shimizu Y. K., Iwamoto A., Hijikata M., Purcell R. H., Yoshikura H. 1992; Evidence forin vitro replication of hepatitis C virus genome in a human T-cell line. Proceedings of the National Academy of SciencesUSA 89:5477–5481
    [Google Scholar]
  49. 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]
  50. Suzuki M. 1989; SPKK, a new nucleic acid-binding unit of protein found in histone. EMBO Journal 8:797–804
    [Google Scholar]
  51. Suzuki R., Ando A., Harada S., Matsuura Y., Saito I., Miyamura T. 1991; Analysis of the HCV genome coding for core protein. The Third International Symposium on Viral Hepatitis and Hepatocellular Carcinoma Taipei:110
    [Google Scholar]
  52. Tadano M., Makino Y., Fukunaga T., Okuno Y., Fukai K. 1989; Detection of dengue 4 virus core protein in the nucleus. I. A monoclonal antibody to dengue 4 virus reacts with the antigen in the nucleus and cytoplasm. Journal of General Virology 70:1409–1415
    [Google Scholar]
  53. 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]
  54. Takebe Y., Seiki M., Fujisawa J., Hoy P., Yokota K., Arai K., Yoshida M., Arai N. 1988; SRa promoter: an efficient and versatile mammalian cDNA expression system composed of the simian virus 40 early promoter and the R-U5 segment of human T- cell leukemia virus type 1 long terminal repeat. Molecular and Cellular Biology 8:466–472
    [Google Scholar]
  55. 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 nucleotide sequences of two isolates derived from an experimentally infected chimpanzee and healthy human carriers. Journal of General Virology 71:3027–3033
    [Google Scholar]
  56. Van der Poel C. L., Cuypers H. T. M., Reesink H. W., Weiner A. J., Quan S., Nello R. D., Boven J. J. P. V., , Winkel I., Mulder-Folkerts D., Exel-Oehilers P. J., Schaasberg W., Leentvaar-Kuypers A., Polito A., Houghton M., Lelie P. N. 1991; Confirmation of hepatitis C virus infection by new fourantigen recombinant immunoblot assay. Lancet 337:317–319
    [Google Scholar]
  57. Zachow K. R., Ostrow R. S., Faras A. J. 1987; Nucleotide sequence and genome organization of human papillomavirus type 5. Virology 158:251–254
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-76-1-53
Loading
/content/journal/jgv/10.1099/0022-1317-76-1-53
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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