Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1995 Dec 19;92(26):12265–12269. doi: 10.1073/pnas.92.26.12265

Functional antagonism between the retinoic acid receptor and the viral transactivator BZLF1 is mediated by protein-protein interactions.

E Pfitzner 1, P Becker 1, A Rolke 1, R Schüle 1
PMCID: PMC40337  PMID: 8618882

Abstract

The Epstein-Barr virus-encoded protein BZLF1 is a member of the basic leucine zipper (bZip) family of transcription factors. Like several other members of the bZip family, transcriptional activity of BZLF1 is modulated by retinoic acid receptors (RARs). We present evidence that the RAR alpha and BZLF1 can reciprocally repress each other's transcriptional activation by a newly discovered mechanism. Analysis of RAR alpha mutants in transfection studies reveals that the DNA binding domain is sufficient for inhibition of BZLF1 activity. Analysis of BZLF1 mutants indicates that both the coiled-coil dimerization domain and a region containing the transcriptional activation domain of BZLF1 are required for transrepression. Coimmunoprecipitation experiments demonstrate physical interactions between RAR alpha and BZLF1 in vivo. Furthermore, glutathione S-transferase-pulldown assays reveal that these protein-protein interactions are mediated by the coiled-coil dimerization domain of BZLF1 and the DNA binding domain of RAR alpha. While RAR alpha is unable to recognize BZLF1 binding sites, the RAR alpha can be tethered to the DNA by forming a heteromeric complex with BZLF1 bound to DNA. Tethering RARs via protein-protein interactions onto promoter DNA suggest a mechanism through which RARs might gain additional levels of transcriptional regulation.

Full text

PDF
12265

Images in this article

12268Fig. 3
on p.12268
12268Fig. 4
on p.12268
12269Fig. 5
on p.12269

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Dedhar S., Rennie P. S., Shago M., Hagesteijn C. Y., Yang H., Filmus J., Hawley R. G., Bruchovsky N., Cheng H., Matusik R. J. Inhibition of nuclear hormone receptor activity by calreticulin. Nature. 1994 Feb 3;367(6462):480–483. doi: 10.1038/367480a0. [DOI] [PubMed] [Google Scholar]
  2. Flemington E., Speck S. H. Autoregulation of Epstein-Barr virus putative lytic switch gene BZLF1. J Virol. 1990 Mar;64(3):1227–1232. doi: 10.1128/jvi.64.3.1227-1232.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Gutsch D. E., Holley-Guthrie E. A., Zhang Q., Stein B., Blanar M. A., Baldwin A. S., Kenney S. C. The bZIP transactivator of Epstein-Barr virus, BZLF1, functionally and physically interacts with the p65 subunit of NF-kappa B. Mol Cell Biol. 1994 Mar;14(3):1939–1948. doi: 10.1128/mcb.14.3.1939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Kliewer S. A., Umesono K., Mangelsdorf D. J., Evans R. M. Retinoid X receptor interacts with nuclear receptors in retinoic acid, thyroid hormone and vitamin D3 signalling. Nature. 1992 Jan 30;355(6359):446–449. doi: 10.1038/355446a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kouzarides T., Packham G., Cook A., Farrell P. J. The BZLF1 protein of EBV has a coiled coil dimerisation domain without a heptad leucine repeat but with homology to the C/EBP leucine zipper. Oncogene. 1991 Feb;6(2):195–204. [PubMed] [Google Scholar]
  6. Lieberman P. Identification of functional targets of the Zta transcriptional activator by formation of stable preinitiation complex intermediates. Mol Cell Biol. 1994 Dec;14(12):8365–8375. doi: 10.1128/mcb.14.12.8365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Mangelsdorf D. J., Umesono K., Kliewer S. A., Borgmeyer U., Ong E. S., Evans R. M. A direct repeat in the cellular retinol-binding protein type II gene confers differential regulation by RXR and RAR. Cell. 1991 Aug 9;66(3):555–561. doi: 10.1016/0092-8674(81)90018-0. [DOI] [PubMed] [Google Scholar]
  8. Mikaélian I., Drouet E., Marechal V., Denoyel G., Nicolas J. C., Sergeant A. The DNA-binding domain of two bZIP transcription factors, the Epstein-Barr virus switch gene product EB1 and Jun, is a bipartite nuclear targeting sequence. J Virol. 1993 Feb;67(2):734–742. doi: 10.1128/jvi.67.2.734-742.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Packham G., Economou A., Rooney C. M., Rowe D. T., Farrell P. J. Structure and function of the Epstein-Barr virus BZLF1 protein. J Virol. 1990 May;64(5):2110–2116. doi: 10.1128/jvi.64.5.2110-2116.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Perlmann T., Rangarajan P. N., Umesono K., Evans R. M. Determinants for selective RAR and TR recognition of direct repeat HREs. Genes Dev. 1993 Jul;7(7B):1411–1422. doi: 10.1101/gad.7.7b.1411. [DOI] [PubMed] [Google Scholar]
  11. Schwarzmann F., Prang N., Reichelt B., Rinkes B., Haist S., Marschall M., Wolf H. Negatively cis-acting elements in the distal part of the promoter of Epstein-Barr virus trans-activator gene BZLF1. J Gen Virol. 1994 Aug;75(Pt 8):1999–2006. doi: 10.1099/0022-1317-75-8-1999. [DOI] [PubMed] [Google Scholar]
  12. Schüle R., Rangarajan P., Kliewer S., Ransone L. J., Bolado J., Yang N., Verma I. M., Evans R. M. Functional antagonism between oncoprotein c-Jun and the glucocorticoid receptor. Cell. 1990 Sep 21;62(6):1217–1226. doi: 10.1016/0092-8674(90)90397-w. [DOI] [PubMed] [Google Scholar]
  13. Schüle R., Rangarajan P., Yang N., Kliewer S., Ransone L. J., Bolado J., Verma I. M., Evans R. M. Retinoic acid is a negative regulator of AP-1-responsive genes. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6092–6096. doi: 10.1073/pnas.88.14.6092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sista N. D., Barry C., Sampson K., Pagano J. Physical and functional interaction of the Epstein-Barr virus BZLF1 transactivator with the retinoic acid receptors RAR alpha and RXR alpha. Nucleic Acids Res. 1995 May 25;23(10):1729–1736. doi: 10.1093/nar/23.10.1729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sista N. D., Pagano J. S., Liao W., Kenney S. Retinoic acid is a negative regulator of the Epstein-Barr virus protein (BZLF1) that mediates disruption of latent infection. Proc Natl Acad Sci U S A. 1993 May 1;90(9):3894–3898. doi: 10.1073/pnas.90.9.3894. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  17. Umesono K., Murakami K. K., Thompson C. C., Evans R. M. Direct repeats as selective response elements for the thyroid hormone, retinoic acid, and vitamin D3 receptors. Cell. 1991 Jun 28;65(7):1255–1266. doi: 10.1016/0092-8674(91)90020-y. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Zhang Q., Gutsch D., Kenney S. Functional and physical interaction between p53 and BZLF1: implications for Epstein-Barr virus latency. Mol Cell Biol. 1994 Mar;14(3):1929–1938. doi: 10.1128/mcb.14.3.1929. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES