1887

Abstract

The Epstein–Barr virus (EBV) BMRF1 gene product is an essential component of the viral DNA polymerase and is absolutely required for lytic virus replication. In addition to its polymerase accessory protein function, we recently demonstrated that BMRF1 is a transactivator, inducing expression of the essential oriLyt promoter, BHLF1. However, the regions of BMRF1 required for transactivation of BHLF1 are unknown. Here we demonstrate that the carboxy-terminal portion of the BMRF1 protein (amino acids 378–404), although not required for DNA binding or polymerase processivity function, is required for transactivator function as well as nuclear localization. Site-directed mutagenesis of this region allowed us to separate the transactivator and nuclear localization motifs of BMRF1. The two DNA-binding domains of BMRF1 are also required for efficient transactivation of the BHLF1 promoter.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-80-1-69
1999-01-01
2024-04-27
Loading full text...

Full text loading...

/deliver/fulltext/jgv/80/1/0800069a.html?itemId=/content/journal/jgv/10.1099/0022-1317-80-1-69&mimeType=html&fmt=ahah

References

  1. Chen L.-W., Lin L.-S., Chang Y.-S., Liu S. T. 1995; Functional analysis of EA-D of Epstein–Barr virus. Virology 211:593–597
    [Google Scholar]
  2. Chiou J. F., Li J. K., Cheng Y. C. 1985; Demonstration of a stimulatory protein for virus-specified DNA polymerase in phorbol estertreated Epstein–Barr virus-carrying cells. Proceedings of the National Academy of Sciences, USA 82:5728–5731
    [Google Scholar]
  3. Cho M. S., Milman G., Hayward S. D. 1985; A second Epstein–Barr virus early antigen gene in BamHI fragment M encodes a 48- to 50- kilodalton nuclear protein. Journal of Virology 56:860–866
    [Google Scholar]
  4. Dingwall C., Laskey R. A. 1991; Nuclear targeting sequences – a consensus?. Trends in Biochemical Sciences 16:478–481
    [Google Scholar]
  5. Fixman E. D., Hayward G. S., Hayward S. D. 1992; Trans-acting requirements for replication of Epstein–Barr virus ori-Lyt. Journal of Virology 66:5030–5039
    [Google Scholar]
  6. Fixman E. D., Hayward G. S., Hayward S. D. 1995; Replication of Epstein–Barr virus oriLyt: lack of a dedicated virally encoded originbinding protein and dependence on Zta in cotransfection assays. Journal of Virology 69:2998–3006
    [Google Scholar]
  7. Gorman C. M., Moffat L. F., Howard B. H. 1982; Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Molecular and Cellular Biology 2:1044–1051
    [Google Scholar]
  8. Hammerschmidt W., Sugden B. 1988; Identification and characterization of oriLyt, a lytic origin of DNA replication of Epstein–Barr virus. Cell 55:427–433
    [Google Scholar]
  9. Kallin B., Sternas L., Saemundssen A. K., Luka J., Jornvall H., Eriksson B., Tao P. Z., Nilsson M. T., Klein G. 1985; Purification of Epstein–Barr virus DNA polymerase from P3HR-1 cells. Journal of Virology 54:561–568
    [Google Scholar]
  10. Kieff E. 1996; Epstein–Barr virus and its replication. In Fields Virology 3rd edn pp 2343–2396 Edited by Fields B. N., Knipe D. M., Howley P. M. Philadelphia: Lippincott–Raven;
    [Google Scholar]
  11. Kiehl A., Dorsky D. I. 1991; Cooperation of EBV DNA polymerase and EA-D (BMRF1) in vitro and colocalization in nuclei of infected cells. Virology 184:330–340
    [Google Scholar]
  12. Kiehl A., Dorsky D. I. 1995; Bipartite DNA binding region of the Epstein–Barr virus BMRF1 product essential for DNA polymerase accessory function. Journal of Virology 69:1669–1677
    [Google Scholar]
  13. Li J. S., Zhou B. S., Dutschman G. E., Grill S. P., Tan R. S., Cheng Y. C. 1987; Association of Epstein–Barr virus early antigen diffuse component and virus-specified DNA polymerase activity. Journal of Virology 61:2947–2949
    [Google Scholar]
  14. Li Q. X., Young L. S., Niedobitek G., Dawson C. W., Birkenbach M., Wang F., Rickinson A. B. 1992; Epstein–Barr virus infection and replication in a human epithelial cell system. Nature 356:347–350
    [Google Scholar]
  15. Oguro M. O., Shimizu N., Ono Y., Takada K. 1987; Both the rightward and the leftward open reading frames within the BamHI M DNA fragment of Epstein–Barr virus act as trans-activators of gene expression. Journal of Virology 61:3310–3313
    [Google Scholar]
  16. Pearson G. R., Vroman B., Chase B., Sculley T., Hummel M., Kieff E. 1983; Identification of polypeptide components of the Epstein–Barr virus early antigen complex with monoclonal antibodies. Journal of Virology 47:193–201
    [Google Scholar]
  17. Rickinson A. B., Kieff E. 1996; Epstein –Barr virus. In Fields Virology 3rd edn pp 2397–2446 Edited by Fields B. N., Knipe D. M., Howley P. M. Philadelphia: Lippincott–Raven;
    [Google Scholar]
  18. Schepers A., Pich D., Hammerschmidt W. 1993a; A transcription factor with homology to the AP-1 family links RNA transcription and DNA replication in the lytic cycle of Epstein–Barr virus. EMBO Journal 12:3921–3929
    [Google Scholar]
  19. Schepers A., Pich D., Mankertz J., Hammerschmidt W. 1993b; cis-acting elements in the lytic origin of DNA replication of Epstein–Barr virus. Journal of Virology 67:4237–4245
    [Google Scholar]
  20. Sixbey J. W., Nedrud J. G., Raab-Traub N., Hanes R. A., Pagano J. S. 1984; Epstein–Barr virus replication in oropharyngeal epithelial cells. New England Journal of Medicine 310:1225–1230
    [Google Scholar]
  21. Tsai C. H., Glaser R. 1991; A comparison of Epstein–Barr virus specific proteins expressed by three Epstein–Barr virus isolates using specific monoclonal antibodies. Intervirology 32:376–382
    [Google Scholar]
  22. Tsurumi T. 1993; Purification and characterization of the DNA binding activity of the Epstein–Barr virus DNA polymerase accessory protein BMRF1 gene products, as expressed in insect cells by using the baculovirus system. Journal of Virology 67:1681–1687
    [Google Scholar]
  23. Tsurumi T., Daikoku T., Kurachi R., Nishiyama Y. 1993; Functional interaction between Epstein–Barr virus DNA polymerase catalytic subunit and its accessory subunit in vitro . Journal of Virology 67:7648–7653
    [Google Scholar]
  24. Zhang Q., Hong Y., Dorsky D., Holley-Guthrie E., Zalani S., Aly Elshiekh N., Kiehl A., Le T., Kenney S. 1996; Functional and physical interactions between the Epstein–Barr virus (EBV) proteins BZLF1 and BMRF1: effects on EBV transcription and lytic replication. Journal of Virology 70:5131–5142
    [Google Scholar]
  25. Zhang Q., Holley-Guthrie E., Ge J.-Q., Dorsky D., Kenney S. 1997; The Epstein–Barr virus (EBV) DNA polymerase accessory protein, BMRF1, activates the essential downstream component of the EBV oriLyt. Virology 23022–34
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-80-1-69
Loading
/content/journal/jgv/10.1099/0022-1317-80-1-69
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