Role of Epstein–Barr virus-encoded latent membrane protein 2A on virus-induced immortalization and virus activation Free

Abstract

To quantitatively evaluate the role of Epstein-Barr virus (EBV)-encoded latent membrane protein 2A (LMP2A) in immortalization of peripheral B-lymphocytes, we used the Akata cell system to generate an EBV recombinant in which the first exon of the LMP2A gene was disrupted. The results indicated that deletion of the LMP2A gene did not affect the immortalization efficiency of EBV in B-lymphocytes. Deletion of the LMP2A gene made EBV-transformed lymphocytes more permissive for virus replication in response to surface immunoglobulin cross-linking. On the other hand Akata cells, in which LMP2A expression was much lower than in EBV-transformed lymphocytes, were equally permissive for virus replication whether they were infected with wild EBV or LMP2A-knockout EBV. The results raise a question as to the role of LMP2A in inhibition of disruption of virus latency , where LMP2A expression has been expected to be low as in Akata cells.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-82-6-1451
2001-06-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/82/6/0821451a.html?itemId=/content/journal/jgv/10.1099/0022-1317-82-6-1451&mimeType=html&fmt=ahah

References

  1. Baer R., Bankier A. T., Biggin M. D., Deininger P. L., Farrell P. J., Gibson T. J., Hutfull G., Hudson G. S., Satchwell S. C., Seguin C., Tuffnell P. S., Barrell B. G. 1984; DNA sequence and expression of the B95-8 Epstein–Barr virus genomes. Nature 310:207–211
    [Google Scholar]
  2. Brielmeier M., Mautner J., Laux G., Hammerschmidt W. 1996; The latent membrane protein 2 gene of Epstein–Barr virus is important for efficient B cell immortalization. Journal of General Virology 77:2807–2818
    [Google Scholar]
  3. Burkhardt A. L., Bolen J. B., Kieff E., Longnecker R. 1992; An Epstein–Barr virus transformation-associated membrane protein interacts with src family tyrosine kinases. Journal of Virology 66:5161–5167
    [Google Scholar]
  4. Chen F., Zou J., di Renzo L., Winberg G., Hu L., Klein E., Klein G., Ernberg I. 1995; A subpopulation of normal B cells latently infected with Epstein–Barr virus resembles Burkitt lymphoma cells in expressing EBNA-1 but not EBNA-2 or LMP1. Journal of Virology 69:3752–3758
    [Google Scholar]
  5. Fruehling S., Lee S., Herrold R., Frech B., Laux G., Kremmer E., Grasser F. A., Longnecker R. 1996; Identification of latent membrane protein 2A (LMP2A) domains essential for the LMP2A dominant-negative effect on B-lymphocyte surface immunoglobulin signal transduction. Journal of Virology 70:6216–6226
    [Google Scholar]
  6. Kieff E. 1996; Epstein–Barr virus and its replication. In Fields Virology pp 2343–2396 Edited by Fields B. N., Knipe D. M., Howley P. M. Philadelphia: Lippincott–Raven;
    [Google Scholar]
  7. Laux G., Perricaudet M., Farrell P. J. 1988; A spliced Epstein–Barr virus gene expressed in immortalized lymphocytes is created by circularization of the linear viral genome. EMBO Journal 7:769–774
    [Google Scholar]
  8. Longnecker R., Durker B., Roberts T. M., Kieff E. 1991; An Epstein–Barr virus protein associated with cell growth transformation interacts with a tyrosine kinase. Journal of Virology 65:3681–3692
    [Google Scholar]
  9. Longnecker R., Miller C. L., Miao X.-Q., Marchini A., Kieff E. 1992; The only domain which distinguishes Epstein–Barr virus latent membrane protein 2A (LMP2A) from LMP2B is dispensable for lymphocyte infection and growth transformation in vitro; LMP2A is therefore nonessential. Journal of Virology 66:6461–6469
    [Google Scholar]
  10. Longnecker R., Miller C. L., Miao X.-Q., Tomkinson B., Kieff E. 1993; The last seven transmembrane and carboxy-terminal cytoplasmic domains of Epstein–Barr virus latent membrane protein 2 (LMP2) are dispensable for lymphocyte infection and growth transformation in vitro. Journal of Virology 67:2006–2013
    [Google Scholar]
  11. Miller C. L., Longnecker R., Kieff E. 1993; Epstein–Barr virus latent membrane protein 2A blocks calcium mobilization in B lymphocytes. Journal of Virology 67:3087–3094
    [Google Scholar]
  12. Miller C. L., Lee J. H., Kieff E., Burkhardt A. L., Bolen J. B., Longnecker R. 1994a; Epstein–Barr virus protein LMP2A regulates reactivation from latency by negatively regulating tyrosine kinases involved in sIg-mediated signal transduction. Infectious Agents and Disease 3:128–136
    [Google Scholar]
  13. Miller C. L., Lee J. H., Kieff E., Longnecker R. 1994b; An integral membrane protein (LMP2) blocks reactivation of Epstein–Barr virus from latency following surface immunoglobulin crosslinking. Proceedings of the National Academy of Sciences, USA 91:772–776
    [Google Scholar]
  14. Miller C. L., Burkhardt A. L., Lee J. H., Stealey B., Longnecker R., Kieff E. 1995; Integral membrane protein 2 of Epstein–Barr virus regulates reactivation from latency through dominant negative effects on protein-tyrosine kinases. Immunity 2:155–166
    [Google Scholar]
  15. Miyashita E. M., Yang B., Babcock G. J., Thorley-Lawson D. A. 1997; Identification of the site of Epstein–Barr virus persistence in vivo as a resting B cells. Journal of Virology 71:4882–4891
    [Google Scholar]
  16. Qu L., Rowe D. 1992; Epstein–Barr virus latent gene expression in uncultured peripheral blood lymphocytes. Journal of Virology 66:3715–3724
    [Google Scholar]
  17. Rickinson A. B., Kieff E. 1996; In Fields Virology . , 3rd edn. pp 2397–2446 Edited by Fields B. N., Knipe D. M., Howley P. M. Philadelphia: Lippincott–Raven;
  18. Sample J., Liebowitz D., Kieff E. 1989; Two related Epstein–Barr virus membrane proteins are encoded by separate genes. Journal of Virology 63:933–937
    [Google Scholar]
  19. Shimizu N., Tanabe-Tochikura A., Kuroiwa Y., Takada K. 1994; Isolation of Epstein–Barr virus (EBV)-negative cell clones from the EBV-positive Burkitt’s lymphoma (BL) line Akata: malignant phenotypes of BL cells are dependent on EBV. Journal of Virology 68:6069–6073
    [Google Scholar]
  20. Shimizu N., Yoshiyama H., Takada K. 1996; Clonal propagation of Epstein–Barr virus (EBV) recombinants in EBV-negative Akata cells. Journal of Virology 70:7260–7263
    [Google Scholar]
  21. Speck P., Kline K. A., Cheresh P., Longnecker R. 1999; Epstein–Barr virus lacking latent membrane protein 2 immortalizes B cells with efficiency indistinguishable from that of wild-type virus. Journal of General Virology 80:2193–2203
    [Google Scholar]
  22. Takada K. 1984; Cross-linking of cell surface immunoglobulins induces Epstein–Barr virus in Burkitt lymphoma lines. International Journal of Cancer 33:27–32
    [Google Scholar]
  23. Takada K., Ono Y. 1989; Synchronous and sequential activation of latently infected Epstein–Barr virus genomes. Journal of Virology 63:445–449
    [Google Scholar]
  24. Takada K., Horinouchi K., Ono Y., Aya T., Osato T., Takahashi M., Hayasaka S. 1991; An Epstein–Barr virus-producer line Akata: establishment of the cell line and analysis of virus genomes. Virus Genes 5:147–156
    [Google Scholar]
  25. Tierney R. J., Steven N., Young L. S., Rickinson A. B. 1994; Epstein–Barr virus latency in blood mononuclear cells: analysis of viral gene transcription during primary infection and in the carrier state. Journal of Virology 68:7374–7385
    [Google Scholar]
  26. Zimber S. U., Kremmer E., Grasser F., Marschall G., Laux G., Bornkamm G. W. 1993; The Epstein–Barr virus nuclear antigen 2 interacts with an EBNA2 responsive cis-element of the terminal protein 1 gene promoter. EMBO Journal 12:167–175
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-82-6-1451
Loading
/content/journal/jgv/10.1099/0022-1317-82-6-1451
Loading

Data & Media loading...

Most cited Most Cited RSS feed