1887

Abstract

The Epstein—Barr virus (EBV)-encoded, nuclear matrix-associated EBNA-5 protein is preferentially localized within distinct nuclear blobs in EBV-immortalized lymphoblastoid cell lines. We have previously found that the same blobs also contain retinoblastoma (Rb) protein. We now show that they contain hsp70 protein as well. Both EBNA-5 and hsp70 translocate to the nucleolus under cell density congestion or after heat shock. Both proteins relocate to their original position upon the reestablishment of normal physiological conditions. EBNA-5 is tightly bound to the nuclear matrix. The translocated EBNA-5 is also tightly associated with matrix structures, as shown by sequential elution-based cell fractionation. The Rb protein does not translocate to the nucleolus. The virally encoded EBNA-1, -2, -3 and -6, and cellular PCNA, snRNP and cyclin E are not affected either. The translocation of EBNA-5 to the nucleolus is not species- or cell type-specific since stress conditions induced the same phenomenon in EBNA-5-transfected human, mouse and rat cells of different tissue origins.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-76-10-2423
1995-10-01
2024-04-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/76/10/JV0760102423.html?itemId=/content/journal/jgv/10.1099/0022-1317-76-10-2423&mimeType=html&fmt=ahah

References

  1. Adachi Y., Copeland T. D., Hatanaka M., Oroszlan S. 1993; Nucleolar targeting signal of Rex protein of human T-cell leukemia virus type I specifically binds to nucleolar shuttle protein B-23. Journal of Biological Chemistry 268:13930–13934
    [Google Scholar]
  2. Beckmann R., Mizzen L., Welch W. 1990; Interaction of Hsp70 with newly synthesized proteins: implications for protein folding and assembly. Science 248:850–854
    [Google Scholar]
  3. Beckmann R., Lovett M., Welch W. 1992; Examining the function and regulation of hsp70 in cells subjected to metabolic stress. Journal of Cell Biology 117:1137–1150
    [Google Scholar]
  4. Brown C. R., Martin R. L., Hansen W. J., Beckmann R. P., Welch W. J. 1993; The constitutive and stress inducible forms of hsp 70 exhibit functional similarities and interact with one another in an ATP-dependent fashion. Journal of Cell Biology 120:1101–1112
    [Google Scholar]
  5. Carrington W. A., Fogarty K. E. 1987; 3–D molecular distribution in living cells by deconvolution of optical sections using light microscopy. In Proceedings of the 13th Annual Northeast Bioengineering Conference pp 108–111 Edited by Foster K.
    [Google Scholar]
  6. Creancier L., Prats H., Zanibellato C., Amalric F., Bugler B. 1993; Determination of the functional domains involved in nucleolar targeting of nucleolin. Molecular Biology of the Cell 4:1239–1250
    [Google Scholar]
  7. Dillner J., Wendel-Hansen V., Kjellström G., Kallin B., Rosen A. 1988; Purification and characterization of Epstein-Barr virus nuclear antigen 2 using monoclonal antipeptide antibodies. International Journal of Cancer 42:721–727
    [Google Scholar]
  8. Elis R. J., van der Vies H. 1991; Molecular chaperons. Annual Review of Biochemistry 60:321–347
    [Google Scholar]
  9. Fankhauser C., Izaurralde E., Adachi Y., Wingfield P., Laemmli U. K. 1991; Specific complex of human immunodeficiency virus type 1 Rev and nucleolar B23 proteins: dissociation by the Rev response element. Molecular and Cellular Biology 11:2567–2575
    [Google Scholar]
  10. Fay F., Carrington W., Fogarty K. 1989; Three-dimensional molecular distribution in single cells analysed using the digital imaging microscope. Journal of Microscopy 153:133–149
    [Google Scholar]
  11. Finke J., Rowe M., Kallin B., Ernberg I., Rosen A., Dillner J., Klein G. 1987; Monoclonal and polyclonal antibodies against Epstein-Barr virus nuclear antigen 5 (EBNA-5) detect multiple protein species in Burkitfs lymphoma and Iymphoblastoid cell lines. Journal of Virology 61:3870–3878
    [Google Scholar]
  12. Frydman J., Nimmesgern E., Ohtsuka K., Hartl F. 1994; Folding of nascent polypeptide chains in a high molecular mass assembly with molecular chaperones. Nature 370:111–117
    [Google Scholar]
  13. Hattori H., Kaneda T., Lokeshwar B., Laszlo A., Ohtsuka K. 1993; A stress-inducible 40 kDa protein (hsp40): purification by modified two-dimensional gel electrophoresis and co-localization with hsp70 (p73) in heat-shocked HeLa cells. Journal of Cell Science 104:629–638
    [Google Scholar]
  14. Hayashi Y., Tohnai I., Kaneda T., Kobayashi T., Ohtsuka K. 1991; Translocation of hsp-70 and protein synthesis during continuous heating at mild temperatures in HeLa cells. Radiation Research 125:80–88
    [Google Scholar]
  15. Jiang W. Q., Szekely L., Wendel-Hansen V., Ringertz N., Klein G., Rosen A. 1991a; Co-localization of the retinoblastoma protein and the Epstein–Barr virus-encoded nuclear antigen EBNA-5. Experimental Cell Research 197:314–318
    [Google Scholar]
  16. Jiang W. Q., Wendel-Hansen V., Lundkvist A., Ringertz N., Klein G., Ros N. A. 1991b; Intranuclear distribution of Epstein-Barr virus-encoded nuclear antigens EBNA-1, -2, -3 and -5. Journal of Cell Science 99:497–502
    [Google Scholar]
  17. Liu Y. C., Hayashi Y., Tohnai I., Kaneda T., Ohtsuka K. 1992; Effects of continuous heating at mild temperatures on the translocation of hsp70 and protein synthesis in NRK cells. Journal of Radiation Research 33:199–210
    [Google Scholar]
  18. Mannick J. B., Cohen J. I., Birkenbach M., Marchini A., Kieff E. 1991; The Epstein-Barr virus nuclear protein encoded by the leader of the EBNA RNAs is important in B-lymphocyte transformation. Journal of Virology 65:6826–6837
    [Google Scholar]
  19. Ohtsuka K., Utsumi K. R., Kaneda T., Hattori H. 1993; Effect of ATP on the release of hsp 70 and hsp 40 from the nucleolus in heat-shocked HeLa cells. Experimental Cell Research 209:357–366
    [Google Scholar]
  20. Rikkonen M., Peranen J., Kääriäinen L. 1992; Nuclear and nucleolar targeting signals of Semliki Forest virus nonstructural protein nsP2. Virology 189:462–473
    [Google Scholar]
  21. Ringertz N. R., Carlsson S. A., Ege T., Bolund L. 1971; Detection of human and chick nuclear antigens in chick erythrocyte nuclei during reactivation in heterokaryons with HeLa cells. Proceedings of the National Academy of Sciences, USA 68:3228–3232
    [Google Scholar]
  22. Rose K. M., Stetler D. A., Jacobs S. T. 1981; Protein kinase activity of RNA polymerase I purified from a rat hepatoma: probable function of Mr 42000 and 26000 polypeptides. Proceedings of the National Academy of Sciences, USA 78:2833–2837
    [Google Scholar]
  23. Schmidt-Zachmann M., Nigg E. 1993; Protein localization to the nucleolus: a search for targeting domains in nucleolin. Journal of Cell Science 105:799–806
    [Google Scholar]
  24. Sinclair A. J., Palmero I., Peters G., Farrell P. J. 1994; EBNA-2 and EBNA-LP cooperate to cause G0 to G1 transition during immortalization of resting human B lymphocytes by Epstein–Barr virus. EMBO Journal 13:3321–3328
    [Google Scholar]
  25. Sjöberg G., Jiang W.-Q., Ringertz N., Lendahl U., Sejersen T. 1994; Colocalization of nestin and vimentin/desmin in skeletal muscle cells demonstrated by three-dimensional fluorescence digital imaging microscopy. Experimental Cell Research 214:447–458
    [Google Scholar]
  26. Szekely L., Uzvolgyi E., Jiang W. Q., Durko M., Wiman K. G., Klein G., Sumegi J. 1991; Subcellular localization of the retinoblastoma protein. Cell Growth and Differentiation 2:287–295
    [Google Scholar]
  27. Szekely L., Selivanova G., Magnusson K. P., Klein G., Wiman K. G. 1993; EBNA-5, an Epstein-Barr virus-encoded nuclear antigen, binds to the retinoblastoma and p53 proteins. Proceedings of the National Academy of Sciences, USA 90:5455–5459
    [Google Scholar]
  28. Xue Z., Shan X., Lapeyre B., Melese T. 1993; The amino terminus of mammalian nucleolin specifically recognizes SV40 T-antigen type nuclear localization sequences. European Journal of Cell Biology 62:13–21
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-76-10-2423
Loading
/content/journal/jgv/10.1099/0022-1317-76-10-2423
Loading

Data & Media loading...

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