1887

Abstract

Following binding to cell surface sialic acid, entry of influenza viruses into cells is mediated by endocytosis. Productive entry of influenza virus requires the low-pH environment of the late endosome for fusion and release of the virus into the cytoplasm and transport of the virus genome into the nucleus. We investigated novel mechanisms to inhibit influenza virus infection using highly specific inhibitors of protein kinase C. We found that one inhibitor, bisindolylmaleimide I, prevented replication of influenza A virus in a dose-dependent manner when added at the time of infection, but had little specific effect when added 2 h after infection had commenced. Virus yields dropped by more than 3 log units in the presence of micromolar levels of bisindolylmaleimide I. Influenza B virus replication was also inhibited by bisindolylmaleimide at micromolar concentrations. We carried out experiments to determine the point in infection that was blocked by bisindolylmaleimide I, and determined that entry of viral ribonucleoproteins (vRNPs) into the nucleus was prevented. Upon drug washout vRNP nuclear entry resumed, showing that bisindolylmaleimide I is reversible. Bisindolylmaleimide I did not affect virus binding and was apparently not acting as a weak base, because its effects were independent of the pH of the external growth medium. These experiments show that bisindolylmaleimide I blocks replication of different types of influenza virus in a dose-dependent and reversible manner, and that virus entry into the cell is inhibited.

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2000-11-01
2019-10-19
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References

  1. Aballay, A., Stahl, P. D. & Mayorga, L. S. ( 1999; ). Phorbol ester promotes endocytosis by activating a factor involved in endosome fusion. Journal of Cell Science 112, 2549-2557.
    [Google Scholar]
  2. Bradshaw, D., Hill, C., Nixon, J. S. & Wilkinson, S. E. ( 1993; ). Therapeutic potential of protein kinase C inhibitors. Agents and Actions 38, 135-147.[CrossRef]
    [Google Scholar]
  3. Bui, M., Whittaker, G. & Helenius, A. ( 1996; ). Effect of M1 protein and low pH on nuclear transport of influenza virus ribonucleoproteins. Journal of Virology 70, 8391-8401.
    [Google Scholar]
  4. Bui, M., Wills, E., Helenius, A. & Whittaker, G. R. ( 2000; ). The role of influenza virus M1 in nuclear export of viral RNPs. Journal of Virology 74, 1781-1786.[CrossRef]
    [Google Scholar]
  5. Bukrinskaya, A. G., Vorkounova, N. K., Kornilayeva, G. V., Narmanbetova, R. A. & Vorkunova, G. K. ( 1982; ). Influenza virus uncoating in infected cells and effect of rimantadine. Journal of General Virology 60, 49-59.[CrossRef]
    [Google Scholar]
  6. Cirone, M., Angeloni, A., Barile, G., Zompetta, C., Venanzoni, M., Torrisi, M. R., Frati, L. & Faggioni, A. ( 1990; ). Epstein–Barr virus internalization and infectivity are blocked by selective protein kinase C inhibitors. International Journal of Cancer 45, 490-493.[CrossRef]
    [Google Scholar]
  7. Constantinescu, S. N., Cernescu, C. D. & Popescu, L. M. ( 1991; ). Effects of protein kinase C inhibitors on viral entry and infectivity. FEBS Letters 292, 31-33.[CrossRef]
    [Google Scholar]
  8. Cox, N. J. & Kawaoka, Y. (1998). Orthomyxoviruses: influenza. In Topley and Wilson’s Microbiology and Microbial Infections, pp. 385–433. New York: Oxford University Press.
  9. Dimitrov, D. S. ( 1997; ). How do viruses enter cells? The HIV coreceptors teach us a lesson of complexity. Cell 91, 721-730.[CrossRef]
    [Google Scholar]
  10. Ferrari, S. L., Behar, V., Chorev, M., Rosenblatt, M. & Bisello, A. ( 1999; ). Endocytosis of ligand-human parathyroid hormone receptor 1 complexes is protein kinase C-dependent and involves β-arrestin2. Real-time monitoring by fluorescence microscopy. Journal of Biological Chemistry 274, 29968-29975.[CrossRef]
    [Google Scholar]
  11. Garland, L. G., Bonser, R. W. & Thompson, N. T. ( 1987; ). Protein kinase C inhibitors are not selective (letter). Trends in Pharmacological Science 8, 334.
    [Google Scholar]
  12. Hay, A. J. ( 1998; ). The virus genome and its replication. In Textbook of Influenza, pp. 43-53. Edited by K. G. Nicholson, R. G. Webster & A. J. Hay. Oxford:Blackwell Science.
  13. Hay, A. J., Wolstenholme, A. J., Skehel, J. J. & Smith, M. H. ( 1985; ). The molecular basis of the specific anti-influenza action of amantadine. EMBO Journal 4, 3021-3024.
    [Google Scholar]
  14. Hernandez, L. D., Hoffman, L. R., Wolfsberg, T. G. & White, J. M. ( 1996; ). Virus–cell and cell–cell fusion. Annual Review of Cell Development and Biology 12, 627-661.[CrossRef]
    [Google Scholar]
  15. Kato, N. & Eggers, H. J. ( 1969; ). Inhibition of uncoating of fowl plague virus by l-adamantanamine hydrochloride. Virology 37, 632-641.[CrossRef]
    [Google Scholar]
  16. Kielian, M. & Helenius, A. ( 1986; ). Entry of alphaviruses. In The Togaviridae and Flaviviridae, pp. 91-119. Edited by S. Schlesinger & M. J. Schlesinger. New York:Plenum Press.
  17. Kurokawa, M., Ochiai, H., Nakajima, K. & Niwayama, S. ( 1990; ). Inhibitory effect of protein kinase C inhibitor on the replication of influenza type A virus. Journal of General Virology 71, 2149-2155.[CrossRef]
    [Google Scholar]
  18. Lamb, R. A. & Pinto, L. H. ( 1997; ). Do Vpu and Vpr of human immunodeficiency virus type 1 and NB of influenza B virus have ion channel activities in the virus life cycles? Virology 229, 1-11.[CrossRef]
    [Google Scholar]
  19. Marsh, M. & Helenius, A. ( 1989; ). Virus entry into animal cells. Advances in Virus Research 36, 107-151.
    [Google Scholar]
  20. Marsh, M. & Pelchen-Matthews, A. ( 1994; ). The endocytic pathway and virus entry. In Cellular Receptors for Animal Viruses, pp. 215-240. Edited by E. Wimmer. Cold Spring Harbor, NY:Cold Spring Harbor Laboratory.
  21. Martin, K. & Helenius, A. ( 1991; ). Nuclear transport of influenza virus ribonucleoproteins: the viral matrix protein (M1) promotes export and inhibits import. Cell 67, 117-130.[CrossRef]
    [Google Scholar]
  22. Mellor, H. & Parker, P. J. ( 1998; ). The extended protein kinase C family. Biochemical Journal 332, 281-292.
    [Google Scholar]
  23. Pelchen-Matthews, A., Parsons, I. J. & Marsh, M. ( 1993; ). Phorbol ester-induced downregulation of CD4 is a multistep process involving dissociation from p56lck, and increased association with clathrin-coated pits, and endosomal sorting. Journal of Experimental Medicine 178, 1209-1222.[CrossRef]
    [Google Scholar]
  24. Pinto, L. H., Holsinger, L. J. & Lamb, R. A. ( 1992; ). Influenza virus M2 protein has ion channel activity. Cell 69, 517-28.[CrossRef]
    [Google Scholar]
  25. Quick, J., Ware, J. A. & Drieger, P. E. ( 1992; ). The structure and biological activities of the widely used protein kinase inhibitor, H7, differ depending on the commercial source. Biochemical and Biophysical Research Communications 187, 657-663.[CrossRef]
    [Google Scholar]
  26. Roy, A.-M. M., Parker, J. S., Parrish, C. R. & Whittaker, G. R. ( 2000; ). Early stages of influenza virus entry into Mv-1 lung cells: involvement of dynamin. Virology 267, 17-28.[CrossRef]
    [Google Scholar]
  27. Ruigrok, R. W. H. ( 1998; ). Structure of influenza A, B and C viruses. In Textbook of Influenza, pp. 29-42. Edited by K. G. Nicholson, R. G. Webster & A. J. Hay. Oxford:Blackwell Science.
  28. Sanchez, P., de Carcer, G., Sandoval, I. V., Moscat, J. & Diaz-Meco, M. ( 1998; ). Localization of atypical protein kinase C isoforms into lysosome-targeted endosomes through interaction with p62. Molecular and Cellular Biology 18, 3069-3080.
    [Google Scholar]
  29. Schultz-Cherry, S., Dybdahl-Sissoko, N., McGregor, M. & Hinshaw, V. S. ( 1998; ). Mink lung epithelial cells: a unique cell line that supports influenza A and B virus replication. Journal of Clinical Microbiology 36, 3718-3720.
    [Google Scholar]
  30. Signoret, N., Oldridge, J., Pelchen-Matthews, A., Klasse, P. J., Tran, T., Brass, L. F., Rosenkilde, M. M., Schwartz, T. W., Holmes, W., Dallas, W., Luther, M. A., Wells, T. N. C., Hoxie, J. A. & Marsh, M. ( 1997; ). Phorbol esters and SDF-1 induce rapid endocytosis and down modulation of the chemokine receptor CXCR4. Journal of Cell Biology 139, 651-664.[CrossRef]
    [Google Scholar]
  31. Skehel, J. J., Bizebard, T., Bullough, P. A., Hughson, F. M., Knossow, M., Steinhauer, D. A., Wharton, S. A. & Wiley, D. C. ( 1995; ). Membrane fusion by influenza virus. Cold Spring Harbor Symposia on Quantitative Biology LX, 573-580.
    [Google Scholar]
  32. Stegmann, T., Morselt, H. W. M., Scholma, J. & Wilschut, J. ( 1987; ). Fusion of influenza virus in an intracellular acidic compartment measured by fluorescence dequenching. Biochimica et Biophysica Acta 904, 165-170.[CrossRef]
    [Google Scholar]
  33. Stuart-Harris, C. H. ( 1939; ). A neurotropic strain of human influenza virus. Lancet 236, 497-499.
    [Google Scholar]
  34. Sugrue, R. J. & Hay, A. J. ( 1991; ). Structural characteristics of the M2 protein of influenza A viruses: evidence that it forms a tetrameric channel. Virology 180, 617-624.[CrossRef]
    [Google Scholar]
  35. Toker, A. ( 1998; ). Signaling through protein kinase C. Frontiers in Bioscience 3, D1134-D1147.
    [Google Scholar]
  36. Toullec, D., Pianetti, P., Coste, H., Bellevergue, P., Grand-Perret, T., Ajakane, M., Baude, T. V., Boissin, P., Boursier, E. & Loriolle F. and others ( 1991; ). The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C. Journal of Biological Chemistry 266, 15771–15781.
    [Google Scholar]
  37. Vogel, U., Kunerl, M. & Scholtissek, C. ( 1994; ). Influenza A virus late mRNAs are specifically retained in the nucleus in the presence of a methyltransferase or a protein kinase inhibitor. Virology 198, 227-233.[CrossRef]
    [Google Scholar]
  38. White, J., Kartenbeck, J. & Helenius, A. ( 1982; ). Membrane fusion activity of influenza virus. EMBO Journal 1, 217-222.
    [Google Scholar]
  39. Whittaker, G., Kemler, I. & Helenius, A. ( 1995; ). Hyperphosphorylation of mutant influenza virus matrix (M1) protein causes its retention in the nucleus. Journal of Virology 69, 439-445.
    [Google Scholar]
  40. Whittaker, G., Bui, M. & Helenius, A. ( 1996; ). The role of nuclear import and export in influenza virus infection. Trends in Cell Biology 6, 67-71.[CrossRef]
    [Google Scholar]
  41. Yoshimura, A. & Ohnishi, S. ( 1984; ). Uncoating of influenza virus in endosomes. Journal of Virology 51, 497-504.
    [Google Scholar]
  42. Yoshimura, A., Kuroda, K., Kawasaki, K., Yamashina, S., Maeda, T. & Ohnishi, S.-I. ( 1982; ). Infectious cell entry mechanism of influenza virus. Journal of Virology 43, 284-293.
    [Google Scholar]
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