Skip to main content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
J Virol. 1996 Jun; 70(6): 4179–4183.
doi: 10.1128/jvi.70.6.4179-4183.1996
PMCID: PMC190314
PMID: 8648764

EBNA-2 and EBNA-3C extensively and mutually exclusively associate with RBPJkappa in Epstein-Barr virus-transformed B lymphocytes.

E Johannsen, C L Miller, S R Grossman, and E Kieff
Author information Copyright and License information PMC Disclaimer

Abstract

Although genetic and biochemical data indicate that the cell protein RBPJkappa is a mediator of EBNA-2 and EBNA-3C effects on transcriptional regulatory elements, the extent of association of these Epstein-Barr virus nuclear proteins with RBPJkappa in transformed B lymphocytes has not been determined. We now report that most of the EBNA-2 and at least 20% of the EBNA-3C coimmunoprecipitated with RBPJkappa from extracts of transformed B lymphocytes that contained most of the cellular EBNA-2 and EBNA3C. Both proteins are associated preferentially with the smaller of the two RBPJkappa isoforms. EBNA-2-RBPJkappa complexes do not contain EBNA-3C, and EBNA-3C-RBPJkappa complexes do not contain EBNA-2. Although EBNA-2 and EBNA-3C are extensively associated with RBPJkappa, a fraction of RBPJkappa appears to be free of EBNAs after repeated immunoprecipitations with anti-EBNA, Epstein-Barr virus-immune, human antibody. Promoters with RBPJkappa sites in their regulatory elements are likely to be differentially regulated by these RBPJkappa-EBNA-2 and RBPJkappa-EBNA-3 complexes.

Full Text

The Full Text of this article is available as a PDF (372K).

Selected References

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

  • Abbot SD, Rowe M, Cadwallader K, Ricksten A, Gordon J, Wang F, Rymo L, Rickinson AB. Epstein-Barr virus nuclear antigen 2 induces expression of the virus-encoded latent membrane protein. J Virol. 1990 May;64(5):2126–2134. [PMC free article] [PubMed] [Google Scholar]
  • Allday MJ, Farrell PJ. Epstein-Barr virus nuclear antigen EBNA3C/6 expression maintains the level of latent membrane protein 1 in G1-arrested cells. J Virol. 1994 Jun;68(6):3491–3498. [PMC free article] [PubMed] [Google Scholar]
  • Amakawa R, Jing W, Ozawa K, Matsunami N, Hamaguchi Y, Matsuda F, Kawaichi M, Honjo T. Human Jk recombination signal binding protein gene (IGKJRB): comparison with its mouse homologue. Genomics. 1993 Aug;17(2):306–315. [PubMed] [Google Scholar]
  • Artavanis-Tsakonas S, Matsuno K, Fortini ME. Notch signaling. Science. 1995 Apr 14;268(5208):225–232. [PubMed] [Google Scholar]
  • Brou C, Logeat F, Lecourtois M, Vandekerckhove J, Kourilsky P, Schweisguth F, Israël A. Inhibition of the DNA-binding activity of Drosophila suppressor of hairless and of its human homolog, KBF2/RBP-J kappa, by direct protein-protein interaction with Drosophila hairless. Genes Dev. 1994 Oct 15;8(20):2491–2503. [PubMed] [Google Scholar]
  • Cohen JI, Wang F, Kieff E. Epstein-Barr virus nuclear protein 2 mutations define essential domains for transformation and transactivation. J Virol. 1991 May;65(5):2545–2554. [PMC free article] [PubMed] [Google Scholar]
  • Otto H, Marti T, Holz M, Mogi T, Lindau M, Khorana HG, Heyn MP. Aspartic acid-96 is the internal proton donor in the reprotonation of the Schiff base of bacteriorhodopsin. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9228–9232. [PMC free article] [PubMed] [Google Scholar]
  • Couso JP, Martinez Arias A. Notch is required for wingless signaling in the epidermis of Drosophila. Cell. 1994 Oct 21;79(2):259–272. [PubMed] [Google Scholar]
  • Dou S, Zeng X, Cortes P, Erdjument-Bromage H, Tempst P, Honjo T, Vales LD. The recombination signal sequence-binding protein RBP-2N functions as a transcriptional repressor. Mol Cell Biol. 1994 May;14(5):3310–3319. [PMC free article] [PubMed] [Google Scholar]
  • Ellisen LW, Bird J, West DC, Soreng AL, Reynolds TC, Smith SD, Sklar J. TAN-1, the human homolog of the Drosophila notch gene, is broken by chromosomal translocations in T lymphoblastic neoplasms. Cell. 1991 Aug 23;66(4):649–661. [PubMed] [Google Scholar]
  • Finke J, Rowe M, Kallin B, Ernberg I, Rosén A, Dillner J, Klein G. Monoclonal and polyclonal antibodies against Epstein-Barr virus nuclear antigen 5 (EBNA-5) detect multiple protein species in Burkitt's lymphoma and lymphoblastoid cell lines. J Virol. 1987 Dec;61(12):3870–3878. [PMC free article] [PubMed] [Google Scholar]
  • Fortini ME, Artavanis-Tsakonas S. The suppressor of hairless protein participates in notch receptor signaling. Cell. 1994 Oct 21;79(2):273–282. [PubMed] [Google Scholar]
  • Furukawa T, Maruyama S, Kawaichi M, Honjo T. The Drosophila homolog of the immunoglobulin recombination signal-binding protein regulates peripheral nervous system development. Cell. 1992 Jun 26;69(7):1191–1197. [PubMed] [Google Scholar]
  • Goodbourn S. Signal transduction. Notch takes a short cut. Nature. 1995 Sep 28;377(6547):288–289. [PubMed] [Google Scholar]
  • Hamaguchi Y, Yamamoto Y, Iwanari H, Maruyama S, Furukawa T, Matsunami N, Honjo T. Biochemical and immunological characterization of the DNA binding protein (RBP-J kappa) to mouse J kappa recombination signal sequence. J Biochem. 1992 Sep;112(3):314–320. [PubMed] [Google Scholar]
  • Hammerschmidt W, Sugden B. Genetic analysis of immortalizing functions of Epstein-Barr virus in human B lymphocytes. Nature. 1989 Aug 3;340(6232):393–397. [PubMed] [Google Scholar]
  • Henkel T, Ling PD, Hayward SD, Peterson MG. Mediation of Epstein-Barr virus EBNA2 transactivation by recombination signal-binding protein J kappa. Science. 1994 Jul 1;265(5168):92–95. [PubMed] [Google Scholar]
  • Hennessy K, Kieff E. A second nuclear protein is encoded by Epstein-Barr virus in latent infection. Science. 1985 Mar 8;227(4691):1238–1240. [PubMed] [Google Scholar]
  • Jarriault S, Brou C, Logeat F, Schroeter EH, Kopan R, Israel A. Signalling downstream of activated mammalian Notch. Nature. 1995 Sep 28;377(6547):355–358. [PubMed] [Google Scholar]
  • Johannsen E, Koh E, Mosialos G, Tong X, Kieff E, Grossman SR. Epstein-Barr virus nuclear protein 2 transactivation of the latent membrane protein 1 promoter is mediated by J kappa and PU.1. J Virol. 1995 Jan;69(1):253–262. [PMC free article] [PubMed] [Google Scholar]
  • Kempkes B, Pich D, Zeidler R, Sugden B, Hammerschmidt W. Immortalization of human B lymphocytes by a plasmid containing 71 kilobase pairs of Epstein-Barr virus DNA. J Virol. 1995 Jan;69(1):231–238. [PMC free article] [PubMed] [Google Scholar]
  • Laux G, Adam B, Strobl LJ, Moreau-Gachelin F. The Spi-1/PU.1 and Spi-B ets family transcription factors and the recombination signal binding protein RBP-J kappa interact with an Epstein-Barr virus nuclear antigen 2 responsive cis-element. EMBO J. 1994 Dec 1;13(23):5624–5632. [PMC free article] [PubMed] [Google Scholar]
  • Lecourtois M, Schweisguth F. The neurogenic suppressor of hairless DNA-binding protein mediates the transcriptional activation of the enhancer of split complex genes triggered by Notch signaling. Genes Dev. 1995 Nov 1;9(21):2598–2608. [PubMed] [Google Scholar]
  • Le Roux A, Kerdiles B, Walls D, Dedieu JF, Perricaudet M. The Epstein-Barr virus determined nuclear antigens EBNA-3A, -3B, and -3C repress EBNA-2-mediated transactivation of the viral terminal protein 1 gene promoter. Virology. 1994 Dec;205(2):596–602. [PubMed] [Google Scholar]
  • Ling PD, Rawlins DR, Hayward SD. The Epstein-Barr virus immortalizing protein EBNA-2 is targeted to DNA by a cellular enhancer-binding protein. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9237–9241. [PMC free article] [PubMed] [Google Scholar]
  • Marshall D, Sample C. Epstein-Barr virus nuclear antigen 3C is a transcriptional regulator. J Virol. 1995 Jun;69(6):3624–3630. [PMC free article] [PubMed] [Google Scholar]
  • Matsunami N, Hamaguchi Y, Yamamoto Y, Kuze K, Kangawa K, Matsuo H, Kawaichi M, Honjo T. A protein binding to the J kappa recombination sequence of immunoglobulin genes contains a sequence related to the integrase motif. Nature. 1989 Dec 21;342(6252):934–937. [PubMed] [Google Scholar]
  • Maunders MJ, Petti L, Rowe M. Precipitation of the Epstein-Barr virus protein EBNA 2 by an EBNA 3c-specific monoclonal antibody. J Gen Virol. 1994 Apr;75(Pt 4):769–778. [PubMed] [Google Scholar]
  • Petti L, Sample C, Kieff E. Subnuclear localization and phosphorylation of Epstein-Barr virus latent infection nuclear proteins. Virology. 1990 Jun;176(2):563–574. [PubMed] [Google Scholar]
  • Robertson ES, Lin J, Kieff E. The amino-terminal domains of Epstein-Barr virus nuclear proteins 3A, 3B, and 3C interact with RBPJ(kappa). J Virol. 1996 May;70(5):3068–3074. [PMC free article] [PubMed] [Google Scholar]
  • Robertson ES, Grossman S, Johannsen E, Miller C, Lin J, Tomkinson B, Kieff E. Epstein-Barr virus nuclear protein 3C modulates transcription through interaction with the sequence-specific DNA-binding protein J kappa. J Virol. 1995 May;69(5):3108–3116. [PMC free article] [PubMed] [Google Scholar]
  • Schweisguth F, Posakony JW. Suppressor of Hairless, the Drosophila homolog of the mouse recombination signal-binding protein gene, controls sensory organ cell fates. Cell. 1992 Jun 26;69(7):1199–1212. [PubMed] [Google Scholar]
  • Silins SL, Sculley TB. Modulation of vimentin, the CD40 activation antigen and Burkitt's lymphoma antigen (CD77) by the Epstein-Barr virus nuclear antigen EBNA-4. Virology. 1994 Jul;202(1):16–24. [PubMed] [Google Scholar]
  • Tomkinson B, Kieff E. Use of second-site homologous recombination to demonstrate that Epstein-Barr virus nuclear protein 3B is not important for lymphocyte infection or growth transformation in vitro. J Virol. 1992 May;66(5):2893–2903. [PMC free article] [PubMed] [Google Scholar]
  • Tomkinson B, Robertson E, Kieff E. Epstein-Barr virus nuclear proteins EBNA-3A and EBNA-3C are essential for B-lymphocyte growth transformation. J Virol. 1993 Apr;67(4):2014–2025. [PMC free article] [PubMed] [Google Scholar]
  • Tong X, Drapkin R, Reinberg D, Kieff E. The 62- and 80-kDa subunits of transcription factor IIH mediate the interaction with Epstein-Barr virus nuclear protein 2. Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3259–3263. [PMC free article] [PubMed] [Google Scholar]
  • Tong X, Drapkin R, Yalamanchili R, Mosialos G, Kieff E. The Epstein-Barr virus nuclear protein 2 acidic domain forms a complex with a novel cellular coactivator that can interact with TFIIE. Mol Cell Biol. 1995 Sep;15(9):4735–4744. [PMC free article] [PubMed] [Google Scholar]
  • Tong X, Wang F, Thut CJ, Kieff E. The Epstein-Barr virus nuclear protein 2 acidic domain can interact with TFIIB, TAF40, and RPA70 but not with TATA-binding protein. J Virol. 1995 Jan;69(1):585–588. [PMC free article] [PubMed] [Google Scholar]
  • Wang F, Gregory C, Sample C, Rowe M, Liebowitz D, Murray R, Rickinson A, Kieff E. Epstein-Barr virus latent membrane protein (LMP1) and nuclear proteins 2 and 3C are effectors of phenotypic changes in B lymphocytes: EBNA-2 and LMP1 cooperatively induce CD23. J Virol. 1990 May;64(5):2309–2318. [PMC free article] [PubMed] [Google Scholar]
  • Wang F, Tsang SF, Kurilla MG, Cohen JI, Kieff E. Epstein-Barr virus nuclear antigen 2 transactivates latent membrane protein LMP1. J Virol. 1990 Jul;64(7):3407–3416. [PMC free article] [PubMed] [Google Scholar]
  • Yalamanchili R, Harada S, Kieff E. The N-terminal half of EBNA2, except for seven prolines, is not essential for primary B-lymphocyte growth transformation. J Virol. 1996 Apr;70(4):2468–2473. [PMC free article] [PubMed] [Google Scholar]
  • Yalamanchili R, Tong X, Grossman S, Johannsen E, Mosialos G, Kieff E. Genetic and biochemical evidence that EBNA 2 interaction with a 63-kDa cellular GTG-binding protein is essential for B lymphocyte growth transformation by EBV. Virology. 1994 Nov 1;204(2):634–641. [PubMed] [Google Scholar]
  • Young L, Alfieri C, Hennessy K, Evans H, O'Hara C, Anderson KC, Ritz J, Shapiro RS, Rickinson A, Kieff E, et al. Expression of Epstein-Barr virus transformation-associated genes in tissues of patients with EBV lymphoproliferative disease. N Engl J Med. 1989 Oct 19;321(16):1080–1085. [PubMed] [Google Scholar]
  • Zimber-Strobl U, Strobl LJ, Meitinger C, Hinrichs R, Sakai T, Furukawa T, Honjo T, Bornkamm GW. Epstein-Barr virus nuclear antigen 2 exerts its transactivating function through interaction with recombination signal binding protein RBP-J kappa, the homologue of Drosophila Suppressor of Hairless. EMBO J. 1994 Oct 17;13(20):4973–4982. [PMC free article] [PubMed] [Google Scholar]
Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)
-