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J Virol. 1996 Sep; 70(9): 5909–5915.
doi: 10.1128/jvi.70.9.5909-5915.1996
PMCID: PMC190609
PMID: 8709211

Epstein-Barr virus EBNA3A and EBNA3C proteins both repress RBP-J kappa-EBNA2-activated transcription by inhibiting the binding of RBP-J kappa to DNA.

L Waltzer, M Perricaudet, A Sergeant, and E Manet
Author information Copyright and License information PMC Disclaimer

Abstract

Following infection by Epstein-Barr virus (EBV), the production of viral nuclear proteins EBNA1, EBNA2, EBNA3A, and EBNA3C and the viral membrane protein LMP1 is essential for the permanent proliferation of primary B lymphocytes to occur. Among these, the transcription factor EBNA2 is central to the immortalizing process, since it activates not only the transcription of all the EBNA proteins and LMP1, TP1, and TP2 but also certain cellular genes. EBNA2 is targeted to its DNA-responsive elements through direct interaction with the DNA-binding cellular repressor RBP-J kappa. In a transient-expression assay, the EBNA2-activated transcription was found to be downregulated by EBNA3A, EBNA3B, and EBNA3C. However, since it has been reported that EBNA3C, but not EBNA3A, directly contacts RBP-J kappa in vitro, these proteins appear to repress through different mechanisms. Here, we report for the first time that EBNA3A and EBNA3C both stably interact with RBP-J kappa and most probably repress EBNA2-activated transcription by destabilizing the binding of RBP-J kappa to DNA.

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Selected References

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  • 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]
  • Ace CI, Dalrymple MA, Ramsay FH, Preston VG, Preston CM. Mutational analysis of the herpes simplex virus type 1 trans-inducing factor Vmw65. J Gen Virol. 1988 Oct;69(Pt 10):2595–2605. [PubMed] [Google Scholar]
  • Alfieri C, Birkenbach M, Kieff E. Early events in Epstein-Barr virus infection of human B lymphocytes. Virology. 1991 Apr;181(2):595–608. [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]
  • 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]
  • Calender A, Billaud M, Aubry JP, Banchereau J, Vuillaume M, Lenoir GM. Epstein-Barr virus (EBV) induces expression of B-cell activation markers on in vitro infection of EBV-negative B-lymphoma cells. Proc Natl Acad Sci U S A. 1987 Nov;84(22):8060–8064. [PMC free article] [PubMed] [Google Scholar]
  • Cohen JI, Wang F, Mannick J, Kieff E. Epstein-Barr virus nuclear protein 2 is a key determinant of lymphocyte transformation. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9558–9562. [PMC free article] [PubMed] [Google Scholar]
  • Cordier M, Calender A, Billaud M, Zimber U, Rousselet G, Pavlish O, Banchereau J, Tursz T, Bornkamm G, Lenoir GM. Stable transfection of Epstein-Barr virus (EBV) nuclear antigen 2 in lymphoma cells containing the EBV P3HR1 genome induces expression of B-cell activation molecules CD21 and CD23. J Virol. 1990 Mar;64(3):1002–1013. [PMC free article] [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]
  • Fåhraeus R, Jansson A, Ricksten A, Sjöblom A, Rymo L. Epstein-Barr virus-encoded nuclear antigen 2 activates the viral latent membrane protein promoter by modulating the activity of a negative regulatory element. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7390–7394. [PMC free article] [PubMed] [Google Scholar]
  • Grossman SR, Johannsen E, Tong X, Yalamanchili R, Kieff E. The Epstein-Barr virus nuclear antigen 2 transactivator is directed to response elements by the J kappa recombination signal binding protein. Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7568–7572. [PMC free article] [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]
  • Hammerschmidt W, Sugden B, Baichwal VR. The transforming domain alone of the latent membrane protein of Epstein-Barr virus is toxic to cells when expressed at high levels. J Virol. 1989 Jun;63(6):2469–2475. [PMC free article] [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]
  • Hsieh JJ, Hayward SD. Masking of the CBF1/RBPJ kappa transcriptional repression domain by Epstein-Barr virus EBNA2. Science. 1995 Apr 28;268(5210):560–563. [PubMed] [Google Scholar]
  • Jin XW, Speck SH. Identification of critical cis elements involved in mediating Epstein-Barr virus nuclear antigen 2-dependent activity of an enhancer located upstream of the viral BamHI C promoter. J Virol. 1992 May;66(5):2846–2852. [PMC free article] [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]
  • Kaye KM, Izumi KM, Kieff E. Epstein-Barr virus latent membrane protein 1 is essential for B-lymphocyte growth transformation. Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):9150–9154. [PMC free article] [PubMed] [Google Scholar]
  • Kempkes B, Spitkovsky D, Jansen-Dürr P, Ellwart JW, Kremmer E, Delecluse HJ, Rottenberger C, Bornkamm GW, Hammerschmidt W. B-cell proliferation and induction of early G1-regulating proteins by Epstein-Barr virus mutants conditional for EBNA2. EMBO J. 1995 Jan 3;14(1):88–96. [PMC free article] [PubMed] [Google Scholar]
  • Knutson JC. The level of c-fgr RNA is increased by EBNA-2, an Epstein-Barr virus gene required for B-cell immortalization. J Virol. 1990 Jun;64(6):2530–2536. [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]
  • 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]
  • Mannick JB, Cohen JI, Birkenbach M, Marchini A, Kieff E. The Epstein-Barr virus nuclear protein encoded by the leader of the EBNA RNAs is important in B-lymphocyte transformation. J Virol. 1991 Dec;65(12):6826–6837. [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]
  • Martin JM, Veis D, Korsmeyer SJ, Sugden B. Latent membrane protein of Epstein-Barr virus induces cellular phenotypes independently of expression of Bcl-2. J Virol. 1993 Sep;67(9):5269–5278. [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]
  • Rooney CM, Brimmell M, Buschle M, Allan G, Farrell PJ, Kolman JL. Host cell and EBNA-2 regulation of Epstein-Barr virus latent-cycle promoter activity in B lymphocytes. J Virol. 1992 Jan;66(1):496–504. [PMC free article] [PubMed] [Google Scholar]
  • Sinclair AJ, Palmero I, Peters G, Farrell PJ. EBNA-2 and EBNA-LP cooperate to cause G0 to G1 transition during immortalization of resting human B lymphocytes by Epstein-Barr virus. EMBO J. 1994 Jul 15;13(14):3321–3328. [PMC free article] [PubMed] [Google Scholar]
  • Sjöblom A, Jansson A, Yang W, Laín S, Nilsson T, Rymo L. PU box-binding transcription factors and a POU domain protein cooperate in the Epstein-Barr virus (EBV) nuclear antigen 2-induced transactivation of the EBV latent membrane protein 1 promoter. J Gen Virol. 1995 Nov;76(Pt 11):2679–2692. [PubMed] [Google Scholar]
  • Sjöblom A, Nerstedt A, Jansson A, Rymo L. Domains of the Epstein-Barr virus nuclear antigen 2 (EBNA2) involved in the transactivation of the latent membrane protein 1 and the EBNA Cp promoters. J Gen Virol. 1995 Nov;76(Pt 11):2669–2678. [PubMed] [Google Scholar]
  • Sung NS, Kenney S, Gutsch D, Pagano JS. EBNA-2 transactivates a lymphoid-specific enhancer in the BamHI C promoter of Epstein-Barr virus. J Virol. 1991 May;65(5):2164–2169. [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]
  • Waltzer L, Bourillot PY, Sergeant A, Manet E. RBP-J kappa repression activity is mediated by a co-repressor and antagonized by the Epstein-Barr virus transcription factor EBNA2. Nucleic Acids Res. 1995 Dec 25;23(24):4939–4945. [PMC free article] [PubMed] [Google Scholar]
  • Waltzer L, Logeat F, Brou C, Israel A, Sergeant A, Manet E. The human J kappa recombination signal sequence binding protein (RBP-J kappa) targets the Epstein-Barr virus EBNA2 protein to its DNA responsive elements. EMBO J. 1994 Dec 1;13(23):5633–5638. [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]
  • Webster NJ, Green S, Tasset D, Ponglikitmongkol M, Chambon P. The transcriptional activation function located in the hormone-binding domain of the human oestrogen receptor is not encoded in a single exon. EMBO J. 1989 May;8(5):1441–1446. [PMC free article] [PubMed] [Google Scholar]
  • Woisetschlaeger M, Yandava CN, Furmanski LA, Strominger JL, Speck SH. Promoter switching in Epstein-Barr virus during the initial stages of infection of B lymphocytes. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1725–1729. [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]
  • Zimber-Strobl U, Kremmer E, Grässer F, Marschall G, Laux G, Bornkamm GW. The Epstein-Barr virus nuclear antigen 2 interacts with an EBNA2 responsive cis-element of the terminal protein 1 gene promoter. EMBO J. 1993 Jan;12(1):167–175. [PMC free article] [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]
  • Zimber-Strobl U, Suentzenich KO, Laux G, Eick D, Cordier M, Calender A, Billaud M, Lenoir GM, Bornkamm GW. Epstein-Barr virus nuclear antigen 2 activates transcription of the terminal protein gene. J Virol. 1991 Jan;65(1):415–423. [PMC free article] [PubMed] [Google Scholar]
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