1887

Abstract

In dengue virus (DEN) particles, the core protein is a structural protein of the nucleocapsid. The core protein is known to be present in the nucleus of DEN-infected cells but there have been conflicting reports as to whether it is also present in the nucleolus. To clarify this, the intracellular location of the core protein was examined using a monoclonal antibody, 15B11, which was produced in this study. Immunofluorescence staining with this antibody demonstrated that the core protein first appeared in the cytoplasm and then in the nuclei and nucleoli of infected cells. Nuclear localization of the core protein was determined to be independent of other DEN proteins, since recombinant core proteins still entered the nuclei and nucleoli of cells transfected with only the core protein gene. Three putative nuclear localization signal motifs have been predicted to be present on the core protein. Deletion of the first one (KKAR), located at aa 6–9, and mutation of the second one (KKSK), located at aa 73–76, did not eliminate the nuclear localization property of the core protein. The third motif with a bipartite structure, RKeigrmlnilnRRRR, located at aa 85–100, was determined to be responsible for the nuclear localization of the core protein, since the core protein without this motif was located exclusively in the cytoplasm of DEN-infected cells and that this motif mediated nuclear localization of a normally cytoplasmic protein.

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2002-12-01
2024-12-08
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References

  1. Avirutnan P., Malasit P., Seliger B., Bhakdi S., Husmann M. 1998; Dengue virus infection of human endothelial cells leads to chemokine production, complement activation, and apoptosis. Journal of Immunology 161:6338–6346
    [Google Scholar]
  2. Barrett A. D. 1997; Japanese encephalitis and dengue vaccines. Biologicals 25:27–34
    [Google Scholar]
  3. 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]
  4. Burglin T. R., De Robertis E. M. 1987; The nuclear migration signal of Xenopus laevis nucleoplasmin. EMBO Journal 6:2617–2625
    [Google Scholar]
  5. 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]
  6. Chang C. J., Luh H. W., Wang S. H., Lin H. J., Lee S. C., Hu S. T. 2001; The heterogeneous nuclear ribonucleoprotein K (hnRNP K) interacts with dengue virus core protein. DNA and Cell Biology 20:569–577
    [Google Scholar]
  7. Chatterjee S., Javier M., Stochaj U. 1997; In vivo analysis of nuclear protein traffic in mammalian cells. Experimental Cell Research 236:346–350
    [Google Scholar]
  8. Chu M. C., O’Rourke E. J., Trent D. W. 1989; Genetic relatedness among structural protein genes of dengue 1 virus strains. Journal of General Virology 70:1701–1712
    [Google Scholar]
  9. Claussen M., Rudt F., Pieler T. 1999; Functional modules in ribosomal protein L5 for ribonucleoprotein complex formation and nucleocytoplasmic transport. Journal of Biological Chemistry 274:33951–33958
    [Google Scholar]
  10. Couvelard A., Marianneau P., Bedel C., Drouet M. T., Vachon F., Henin D., Deubel V. 1999; Report of a fatal case of dengue infection with hepatitis: demonstration of dengue antigens in hepatocytes and liver apoptosis. Human Pathology 30:1106–1110
    [Google Scholar]
  11. Despres P., Flamand M., Ceccaldi P. E., Deubel V. 1996; Human isolates of dengue type 1 virus induce apoptosis in mouse neuroblastoma cells. Journal of Virology 70:4090–4096
    [Google Scholar]
  12. Dingwall C., Laskey R. A. 1991; Nuclear targeting sequences: a consensus?. Trends in Biochemical Sciences 16:478–481
    [Google Scholar]
  13. Fontes M. R., Teh T., Kobe B. 2000; Structural basis of recognition of monopartite and bipartite nuclear localization sequences by mammalian importin- α. Journal of Molecular Biology 297:1183–1194
    [Google Scholar]
  14. Gubler D. J., Clark G. G. 1995; Dengue/dengue hemorrhagic fever: the emergence of a global health problem. Emerging Infectious Diseases 1:55–57
    [Google Scholar]
  15. Halstead S. B. 1980; Dengue haemorrhagic fever: a public health problem and a field for research. Bulletin of the World Health Organization 58:1–21
    [Google Scholar]
  16. Hsieh T. Y., Matsumoto M., Chou H. C., Schneider R., Hwang S. B., Lee A. S., Lai M. M. 1998; Hepatitis C virus core protein interacts with heterogeneous nuclear ribonucleoprotein K. Journal of Biological Chemistry 273:17651–17659
    [Google Scholar]
  17. Hwang S. B., Lo S. Y., Ou J. H., Lai M. M. 1995; Detection of cellular proteins and viral core protein interacting with the 5’ untranslated region of hepatitis C virus RNA. Journal of Biomedical Science 2:227–236
    [Google Scholar]
  18. Irie K., Mohan P. M., Sasaguri Y., Putnak R., Padmanabhan R. 1989; Sequence analysis of cloned dengue virus type 2 genome (New Guinea-C strain). Gene 75:197–211
    [Google Scholar]
  19. Jan J. T., Chen B. H., Ma S. H., Liu C. I., Tsai H. P., Wu H. C., Jiang S. Y., Yang K. D., Shaio M. F. 2000; Potential dengue virus-triggered apoptotic pathway in human neuroblastoma cells: arachidonic acid, superoxide anion, and NF-κB are sequentially involved. Journal of Virology 74:8680–8691
    [Google Scholar]
  20. Khromykh A. A., Westaway E. G. 1996; RNA binding properties of core protein of the flavivirus Kunjin. Archives of Virology 141:685–699
    [Google Scholar]
  21. Lane R. D., Crissman R. S., Ginn S. 1986; High efficiency fusion procedure for producing monoclonal antibodies against weak immunogens. Methods in Enzymology 121:183–192
    [Google Scholar]
  22. Lin Y. L., Liu C. C., Lei H. Y., Yeh T. M., Lin Y. S., Chen R. M., Liu H. S. 2000; Infection of five human liver cell lines by dengue-2 virus. Journal of Medical Virology 60:425–431
    [Google Scholar]
  23. Liu J. L., Lee L. F., Ye Y., Qian Z., Kung H. J. 1997; Nucleolar and nuclear localization properties of a herpesvirus bZIP oncoprotein, MEQ. Journal of Virology 71:3188–3196
    [Google Scholar]
  24. Makino Y., Tadano M., Anzai T., Ma S. P., Yasuda S., Fukunaga T. 1989; Detection of dengue 4 virus core protein in the nucleus. II. Antibody against dengue 4 core protein produced by a recombinant baculovirus reacts with the antigen in the nucleus. Journal of General Virology 70:1417–1425
    [Google Scholar]
  25. Marianneau P., Cardona A., Edelman L., Deubel V., Despres P. 1997; Dengue virus replication in human hepatoma cells activates NF- κB which in turn induces apoptotic cell death. Journal of Virology 71:3244–3249
    [Google Scholar]
  26. Marianneau P., Steffan A. M., Royer C., Drouet M. T., Jaeck D., Kirn A., Deubel V. 1999; Infection of primary cultures of human Kupffer cells by dengue virus: no viral progeny synthesis, but cytokine production is evident. Journal of Virology 73:5201–5206
    [Google Scholar]
  27. Michelotti E. F., Michelotti G. A., Aronsohn A. I., Levens D. 1996; Heterogeneous nuclear ribonucleoprotein K is a transcription factor. Molecular and Cellular Biology 16:2350–2360
    [Google Scholar]
  28. Osatomi K., Sumiyoshi H. 1990; Complete nucleotide sequence of dengue type 3 virus genome RNA. Virology 176:643–647
    [Google Scholar]
  29. Pai C. Y., Chen H. K., Sheu H. L., Yeh N. H. 1995; Cell-cycle-dependent alterations of a highly phosphorylated nucleolar protein p130 are associated with nucleologenesis. Journal of Cell Science 108:1911–1920
    [Google Scholar]
  30. Prendergast G. C. 1999; Mechanisms of apoptosis by c-Myc. Oncogene 18:2967–2987
    [Google Scholar]
  31. Reed R. C., Louis-Wileman V., Cosmai E. V., Fang S., Jue D. L., Wohlhueter R. M., Hunter R. L., Lal A. A. 1997; Multiple antigen constructs (MACs): induction of sterile immunity against sporozoite stage of rodent malaria parasites, Plasmodium berghei and Plasmodium yoelii. Vaccine 15:482–488
    [Google Scholar]
  32. Richardson W. D., Roberts B. L., Smith A. E. 1986; Nuclear location signals in polyoma virus large-T. Cell 44:77–85
    [Google Scholar]
  33. Rigau-Perez J. G., Clark G. G., Gubler D. J., Reiter P., Sanders E. J., Vorndam A. V. 1998; Dengue and dengue haemorrhagic fever. Lancet 352:971–977
    [Google Scholar]
  34. 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]
  35. Shimoike T., Mimori S., Tani H., Matsuura Y., Miyamura T. 1999; Interaction of hepatitis C virus core protein with viral sense RNA and suppression of its translation. Journal of Virology 73:9718–9725
    [Google Scholar]
  36. Su H. L., Lin Y. L., Yu H. P., Tsao C. H., Chen L. K., Liu Y. T., Liao C. L. 2001; The effect of human bcl-2 and bcl-X genes on dengue virus-induced apoptosis in cultured cells. Virology 282:141–153
    [Google Scholar]
  37. 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]
  38. Westaway E. G., Brinton M. A., Gaidamovich S., Horzinek M. C., Igarashi A., Kaariainen L., Lvov D. K., Porterfield J. S., Russell P. K., Trent D. W. 1985; Flaviviridae. Intervirology24183–192
    [Google Scholar]
  39. WHO 2000; Dengue/dengue haemorrhagic fever: situation in 2000. Weekly Epidemiological Record 75:193–196
    [Google Scholar]
  40. Zhao B., Mackow E., Buckler-White A., Markoff L., Chanock R. M., Lai C. J., Makino Y. 1986; Cloning full-length dengue type 4 viral DNA sequences: analysis of genes coding for structural proteins. Virology 155:77–88
    [Google Scholar]
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