1887

Abstract

Mouse monoclonal antibodies (MAbs) that specifically detect the 127 kDa Pol precursor and the 85 kDa reverse transcriptase/RNase H (RT/RN) or pr127 and the 40 kDa integrase (IN) in immunoblot and immunofluorescence assays (IFA) were used to investigate the subcellular localization of primate foamy virus (PFV) proteins. IFA of cells infected with PFV using the anti-Pol MAbs and rabbit anti-capsid (Gag) serum revealed that both the Gag and Pol proteins are transported into the nucleus. Transfection of cells with eukaryotic expression constructs for pr127, p85 and p40 served to show Gag-independent subcellular localization of Pol proteins. Interestingly, not only the Pol precursor and IN molecules were found to be localized to the nucleus, but also the RT/RN subdomain. It is therefore suggested that PFV cores bear at least three separate nuclear localization signals, one in Gag and two in Pol. The latter appear to be localized to the two Pol subdomains.

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2000-12-01
2019-10-22
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References

  1. Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D., Seidman, J. G., Smith, J. A. & Struhl, K. (editors) (1987). Current Protocols in Molecular Biology. New York: John Wiley.
  2. Bieniasz, P. D., Weiss, R. A. & McClure, M. O. ( 1995; ). Cell cycle dependence of foamy retrovirus infection. Journal of Virology 69, 7295-7299.
    [Google Scholar]
  3. Coligan, J. E., Kruisbeek, A. M., Margulies, D. H., Shevach, E. M. & Strober, W. (1994). Current Protocols in Immunology. New York: John Wiley.
  4. Enssle, J., Jordan, I., Mauer, B. & Rethwilm, A. ( 1996; ). Foamy virus reverse transcriptase is expressed independently from the Gag protein. Proceedings of the National Academy of Sciences, USA 93, 4137-4141.[CrossRef]
    [Google Scholar]
  5. Falcone, V., Leupold, J., Clotten, J., Urbanyi, E., Herchenröder, O., Spatz, W., Volk, B., Böhm, N., Toniolo, A., Neumann-Haefelin, D. & Schweizer, M. ( 1999; ). Sites of simian foamy virus persistence in naturally infected African green monkeys: latent provirus is ubiquitous, whereas viral replication is restricted to the oral mucosa. Virology 257, 7-14.[CrossRef]
    [Google Scholar]
  6. Fischer, N., Enssle, J., Müller, J., Rethwilm, A. & Niewiesk, S. ( 1997; ). Characterization of human foamy virus proteins expressed by recombinant vaccinia viruses. AIDS Research and Human Retroviruses 13, 517-521.[CrossRef]
    [Google Scholar]
  7. Gallay, P., Hope, T., Chin, D. & Trono, D. ( 1997; ). HIV-1 infection of nondividing cells through the recognition of integrase by the importin/karyopherin pathway. Proceedings of the National Academy of Sciences, USA 94, 9825-9830.[CrossRef]
    [Google Scholar]
  8. Garcia-Bustos, J., Heitman, J. & Hall, M. N. ( 1991; ). Nuclear protein localization. Biochimica et Biophysica Acta 1071, 83-101.[CrossRef]
    [Google Scholar]
  9. Hahn, H., Baunach, G., Bräutigam, S., Mergia, A., Neumann-Haefelin, D., Daniel, M. D., McClure, M. O. & Rethwilm, A. ( 1994; ). Reactivity of primate sera to foamy virus Gag and Bet proteins. Journal of General Virology 75, 2635-2644.[CrossRef]
    [Google Scholar]
  10. Harlow, E. & Lane, D. (1988). Antibodies: A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  11. Heinkelein, M., Schmidt, M., Fischer, N., Moebes, A., Lindemann, D., Enssle, J. & Rethwilm, A. ( 1998; ). Characterization of a cis-acting sequence in the Pol region required to transfer human foamy virus vectors. Journal of Virology 72, 6307-6314.
    [Google Scholar]
  12. Heinkelein, M., Pietschmann, T., Jarmy, G., Dressler, M., Imrich, H., Thurow, J., Lindemann, D., Bock, M., Moebes, A., Roy, J., Herchenröder, O. & Rethwilm, A. ( 2000a; ). Efficient intracellular retrotransposition of an exogenous primate retrovirus genome. EMBO Journal 19, 3436-3445.[CrossRef]
    [Google Scholar]
  13. Heinkelein, M., Thurow, J., Dressler, M., Imrich, H., Neumann-Haefelin, D., McClure, M. O. & Rethwilm, A. ( 2000b; ). Complex effects of deletions in the 5’ untranslated region of primate foamy virus on viral gene expression and RNA packaging. Journal of Virology 74, 3141-3148.[CrossRef]
    [Google Scholar]
  14. Higuchi, R. (1990). Recombinant PCR. In PCR Protocols: A Guide to Methods & Applications, pp. 177–183. Edited by M. A. Innis, D. H. Gelfand & T. J. White. San Diego, CA: Academic Press.
  15. Hooks, J. J. & Gibbs, C. J.Jr ( 1975; ). The foamy viruses. Bacteriological Reviews 39, 169-185.
    [Google Scholar]
  16. Jordan, I., Enssle, J., Güttler, E., Mauer, B. & Rethwilm, A. ( 1996; ). Expression of human foamy virus reverse transcriptase involves a spliced pol mRNA. Virology 224, 314-319.[CrossRef]
    [Google Scholar]
  17. Konvalinka, J., Löchelt, M., Zentgraf, H., Flügel, R. M. & Kräusslich, H.-G. ( 1995; ). Active foamy virus proteinase is essential for virus infectivity but not for formation of a Pol polyprotein. Journal of Virology 69, 7264-7268.
    [Google Scholar]
  18. Kukolj, G., Jones, K. S. & Skalka, A. M. ( 1997; ). Subcellular localization of avian sarcoma virus and human immunodeficiency virus type 1 integrases. Journal of Virology 71, 843-847.
    [Google Scholar]
  19. Kukolj, G., Katz, R. A. & Skalka, A. M. ( 1998; ). Characterization of the nuclear localization signal of the avian sarcoma virus integrase. Gene 223, 157-163.[CrossRef]
    [Google Scholar]
  20. Linial, M. L. ( 1999; ). Foamy viruses are unconventional retroviruses. Journal of Virology 73, 1747-1755.
    [Google Scholar]
  21. Linial, M. L. ( 2000; ). Why aren’t foamy viruses pathogenic? Trends in Microbiology 8, 284-289.[CrossRef]
    [Google Scholar]
  22. Meiering, C. D., Comstock, K. E. & Linial, M. L. ( 2000; ). Multiple integrations of human foamy virus in persistently infected human erythroleukemia cells. Journal of Virology 74, 1718-1726.[CrossRef]
    [Google Scholar]
  23. Moebes, A., Enssle, J., Bieniasz, P. D., Heinkelein, M., Lindemann, D., Bock, M., McClure, M. O. & Rethwilm, A. ( 1997; ). Human foamy virus reverse transcription that occurs late in the viral replication cycle. Journal of Virology 71, 7305-7311.
    [Google Scholar]
  24. Morozov, V. A., Copeland, T. D., Nagashima, K., Gonda, M. A. & Oroszlan, S. ( 1997; ). Protein composition and morphology of human foamy virus intracellular cores and extracellular particles. Virology 228, 307-317.[CrossRef]
    [Google Scholar]
  25. Mumm, S. R., Hippenmeyer, P. J. & Grandgenett, D. P. ( 1992; ). Characterization of a stable eukaryotic cell line expressing the Rous sarcoma virus integrase. Virology 189, 500-510.[CrossRef]
    [Google Scholar]
  26. Nassal, M. & Schaller, H. ( 1993; ). Hepatitis B virus replication. Trends in Microbiology 1, 221-228.[CrossRef]
    [Google Scholar]
  27. Netzer, K.-O., Rethwilm, A., Maurer, B. & ter Meulen, V. ( 1990; ). Identification of the major immunogenic structural proteins of human foamy virus. Journal of General Virology 71, 1237-1241.[CrossRef]
    [Google Scholar]
  28. Petit, C., Schwartz, O. & Mammano, F. ( 1999; ). Oligomerization within virions and subcellular localization of human immunodeficiency virus type 1 integrase. Journal of Virology 73, 5079-5088.
    [Google Scholar]
  29. Pfrepper, K.-I., Rackwitz, H.-R., Schnölzer, M., Heid, H., Löchelt, M. & Flügel, R. M. ( 1998; ). Molecular characterization of proteolytic processing of the Pol proteins of human foamy virus reveals novel features of the viral protease. Journal of Virology 72, 7648-7652.
    [Google Scholar]
  30. Pluymers, W., Cherepanov, P., Schols, D., De Clercq, E. & Debyser, Z. ( 1999; ). Nuclear localization of human immunodeficiency virus type 1 integrase expressed as a fusion protein with green fluorescent protein. Virology 258, 327-332.[CrossRef]
    [Google Scholar]
  31. Popov, S., Rexach, M., Zybarth, G., Reiling, N., Lee, M.-A., Ratner, L., Lane, C. M., Moore, M. S., Blobel, G. & Bukrinsky, M. ( 1998; ). Viral protein R regulates nuclear import of the HIV-1 pre-integration complex. EMBO Journal 17, 909-917.[CrossRef]
    [Google Scholar]
  32. Rethwilm, A. ( 1996; ). Unexpected replication pathways of foamy viruses. Journal of Acquired Immune Deficiency Syndromes and Human Retrovirology 13 (Suppl. 1), S248–S253.[CrossRef]
    [Google Scholar]
  33. Saı̈b, A., Puvion-Dutilleul, F., Schmid, M., Périès, J. & de Thé, H. ( 1997; ). Nuclear targeting of incoming human foamy virus Gag proteins involves a centriolar step. Journal of Virology 71, 1155-1161.
    [Google Scholar]
  34. Schliephake, A. W. & Rethwilm, A. ( 1994; ). Nuclear localization of foamy virus Gag precursor protein. Journal of Virology 68, 4946-4954.
    [Google Scholar]
  35. Schmidt, M. & Rethwilm, A. ( 1995; ). Replicating foamy virus-based vectors directing high level expression of foreign genes. Virology 210, 167-178.[CrossRef]
    [Google Scholar]
  36. Schmidt, M., Niewiesk, S., Heeney, J., Aguzzi, A. & Rethwilm, A. ( 1997; ). Mouse model to study the replication of primate foamy viruses. Journal of General Virology 78, 1929-1933.
    [Google Scholar]
  37. Shulman, M., Wilde, C. D. & Köhler, G. ( 1978; ). A better cell line for making hybridomas secreting specific antibodies. Nature 276, 269-270.[CrossRef]
    [Google Scholar]
  38. Yu, S. F., Edelmann, K., Strong, R. K., Moebes, A., Rethwilm, A. & Linial, M. L. ( 1996a; ). The carboxyl terminus of the human foamy virus Gag protein contains separable nucleic acid binding and nuclear transport domains. Journal of Virology 70, 8255-8262.
    [Google Scholar]
  39. Yu, S. F., Baldwin, D. N., Gwynn, S. R., Yendapalli, S. & Linial, M. L. ( 1996b; ). Human foamy virus replication: a pathway distinct from that of retroviruses and hepadnaviruses. Science 271, 1579-1582.[CrossRef]
    [Google Scholar]
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