Abstract
The Cp promoter of Epstein-Barr virus (EBV) directs most transcription of the EBNA genes in lymphoblastoid cell lines. The functions of two control regions in the Cp promoter have been studied by construction of recombinant EBV strains containing specific mutations in these elements. Mutation of the RBP-Jk (CBF1) binding site reduced but did not completely abolish EBNA-2-dependent Cp activity in transient transfection assays. The same mutation in recombinant virus gave only a modest average reduction in Cp function, ranging from full activity to almost no activity in different isolates. Separate deletion of a 262-bp region containing glucocorticoid response elements had little effect in a transient assay but caused a fivefold increase in the steady-state level of Cp RNA in recombinant virus. The results indicate that other elements in addition to the intensively studied RBP-Jk site are important in determining Cp activity in the whole virus. Clonal EBV-infected cell lines expressed RNA from both the Cp and Wp promoters, but the level of Wp RNA did not simply compensate for changes in the level of Cp RNA. The levels of EBNA proteins varied much less than the levels of Cp and Wp RNA, suggesting other types of control in addition to initiation of transcription. A survey of RNAs derived from the internal repeat region of the virus indicated that gene expression from this region of EBV in lymphoblastoid cell lines is accounted for by the known transcripts.
Full Text
The Full Text of this article is available as a PDF (777.2 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Baer R., Bankier A. T., Biggin M. D., Deininger P. L., Farrell P. J., Gibson T. J., Hatfull G., Hudson G. S., Satchwell S. C., Séguin C. DNA sequence and expression of the B95-8 Epstein-Barr virus genome. Nature. 1984 Jul 19;310(5974):207–211. doi: 10.1038/310207a0. [DOI] [PubMed] [Google Scholar]
- Ben-Bassat H., Goldblum N., Mitrani S., Goldblum T., Yoffey J. M., Cohen M. M., Bentwich Z., Ramot B., Klein E., Klein G. Establishment in continuous culture of a new type of lymphocyte from a "Burkitt like" malignant lymphoma (line D.G.-75). Int J Cancer. 1977 Jan;19(1):27–33. doi: 10.1002/ijc.2910190105. [DOI] [PubMed] [Google Scholar]
- Bodescot M., Brison O., Perricaudet M. An Epstein-Barr virus transcription unit is at least 84 kilobases long. Nucleic Acids Res. 1986 Mar 25;14(6):2611–2620. doi: 10.1093/nar/14.6.2611. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bodescot M., Perricaudet M., Farrell P. J. A promoter for the highly spliced EBNA family of RNAs of Epstein-Barr virus. J Virol. 1987 Nov;61(11):3424–3430. doi: 10.1128/jvi.61.11.3424-3430.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cohen J. I., Kieff E. An Epstein-Barr virus nuclear protein 2 domain essential for transformation is a direct transcriptional activator. J Virol. 1991 Nov;65(11):5880–5885. doi: 10.1128/jvi.65.11.5880-5885.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dou S., Zeng X., Cortes P., Erdjument-Bromage H., Tempst P., Honjo T., Vales L. D. The recombination signal sequence-binding protein RBP-2N functions as a transcriptional repressor. Mol Cell Biol. 1994 May;14(5):3310–3319. doi: 10.1128/mcb.14.5.3310. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Evans T. J., Jacquemin M. G., Farrell P. J. Efficient EBV superinfection of group I Burkitt's lymphoma cells distinguishes requirements for expression of the Cp viral promoter and can activate the EBV productive cycle. Virology. 1995 Feb 1;206(2):866–877. doi: 10.1006/viro.1995.1009. [DOI] [PubMed] [Google Scholar]
- Farrell P. J., Allan G. J., Shanahan F., Vousden K. H., Crook T. p53 is frequently mutated in Burkitt's lymphoma cell lines. EMBO J. 1991 Oct;10(10):2879–2887. doi: 10.1002/j.1460-2075.1991.tb07837.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [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. doi: 10.1128/jvi.61.12.3870-3878.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Furukawa T., Kawaichi M., Matsunami N., Ryo H., Nishida Y., Honjo T. The Drosophila RBP-J kappa gene encodes the binding protein for the immunoglobulin J kappa recombination signal sequence. J Biol Chem. 1991 Dec 5;266(34):23334–23340. [PubMed] [Google Scholar]
- Fåhraeus R., Jansson A., Sjöblom A., Nilsson T., Klein G., Rymo L. Cell phenotype-dependent control of Epstein-Barr virus latent membrane protein 1 gene regulatory sequences. Virology. 1993 Jul;195(1):71–80. doi: 10.1006/viro.1993.1347. [DOI] [PubMed] [Google Scholar]
- Goodbourn S. Signal transduction. Notch takes a short cut. Nature. 1995 Sep 28;377(6547):288–289. doi: 10.1038/377288a0. [DOI] [PubMed] [Google Scholar]
- Grossman S. R., 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. doi: 10.1073/pnas.91.16.7568. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gunning P., Ponte P., Okayama H., Engel J., Blau H., Kedes L. Isolation and characterization of full-length cDNA clones for human alpha-, beta-, and gamma-actin mRNAs: skeletal but not cytoplasmic actins have an amino-terminal cysteine that is subsequently removed. Mol Cell Biol. 1983 May;3(5):787–795. doi: 10.1128/mcb.3.5.787. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Henkel T., Ling P. D., Hayward S. D., Peterson M. G. Mediation of Epstein-Barr virus EBNA2 transactivation by recombination signal-binding protein J kappa. Science. 1994 Jul 1;265(5168):92–95. doi: 10.1126/science.8016657. [DOI] [PubMed] [Google Scholar]
- Hinuma Y., Grace J. T., Jr Cloning of immunoglobulin-producing human leukemic and lymphoma cells in long-term cultures. Proc Soc Exp Biol Med. 1967 Jan;124(1):107–111. doi: 10.3181/00379727-124-31677. [DOI] [PubMed] [Google Scholar]
- Hsieh J. J., Hayward S. D. Masking of the CBF1/RBPJ kappa transcriptional repression domain by Epstein-Barr virus EBNA2. Science. 1995 Apr 28;268(5210):560–563. doi: 10.1126/science.7725102. [DOI] [PubMed] [Google Scholar]
- Hurley E. A., Klaman L. D., Agger S., Lawrence J. B., Thorley-Lawson D. A. The prototypical Epstein-Barr virus-transformed lymphoblastoid cell line IB4 is an unusual variant containing integrated but no episomal viral DNA. J Virol. 1991 Jul;65(7):3958–3963. doi: 10.1128/jvi.65.7.3958-3963.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jin X. W., Speck S. H. 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. doi: 10.1128/jvi.66.5.2846-2852.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Johannsen E., Koh E., Mosialos G., Tong X., Kieff E., Grossman S. R. 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. doi: 10.1128/jvi.69.1.253-262.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kempkes B., Spitkovsky D., Jansen-Dürr P., Ellwart J. W., Kremmer E., Delecluse H. J., Rottenberger C., Bornkamm G. W., 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. doi: 10.1002/j.1460-2075.1995.tb06978.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- King W., Thomas-Powell A. L., Raab-Traub N., Hawke M., Kieff E. Epstein-Barr virus RNA. V. Viral RNA in a restringently infected, growth-transformed cell line. J Virol. 1980 Nov;36(2):506–518. doi: 10.1128/jvi.36.2.506-518.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knott V., Rees D. J., Cheng Z., Brownlee G. G. Randomly picked cosmid clones overlap the pyrB and oriC gap in the physical map of the E. coli chromosome. Nucleic Acids Res. 1988 Mar 25;16(6):2601–2612. doi: 10.1093/nar/16.6.2601. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kupfer S. R., Summers W. C. Identification of a glucocorticoid-responsive element in Epstein-Barr virus. J Virol. 1990 May;64(5):1984–1990. doi: 10.1128/jvi.64.5.1984-1990.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ling P. D., Hsieh J. J., Ruf I. K., Rawlins D. R., Hayward S. D. EBNA-2 upregulation of Epstein-Barr virus latency promoters and the cellular CD23 promoter utilizes a common targeting intermediate, CBF1. J Virol. 1994 Sep;68(9):5375–5383. doi: 10.1128/jvi.68.9.5375-5383.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ling P. D., Rawlins D. R., Hayward S. D. 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. doi: 10.1073/pnas.90.20.9237. [DOI] [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. doi: 10.1038/342934a0. [DOI] [PubMed] [Google Scholar]
- Meitinger C., Strobl L. J., Marschall G., Bornkamm G. W., Zimber-Strobl U. Crucial sequences within the Epstein-Barr virus TP1 promoter for EBNA2-mediated transactivation and interaction of EBNA2 with its responsive element. J Virol. 1994 Nov;68(11):7497–7506. doi: 10.1128/jvi.68.11.7497-7506.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miller G., Lipman M. Release of infectious Epstein-Barr virus by transformed marmoset leukocytes. Proc Natl Acad Sci U S A. 1973 Jan;70(1):190–194. doi: 10.1073/pnas.70.1.190. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morgenstern J. P., Land H. A series of mammalian expression vectors and characterisation of their expression of a reporter gene in stably and transiently transfected cells. Nucleic Acids Res. 1990 Feb 25;18(4):1068–1068. doi: 10.1093/nar/18.4.1068. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rabson M., Gradoville L., Heston L., Miller G. Non-immortalizing P3J-HR-1 Epstein-Barr virus: a deletion mutant of its transforming parent, Jijoye. J Virol. 1982 Dec;44(3):834–844. doi: 10.1128/jvi.44.3.834-844.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rooney C. M., Brimmell M., Buschle M., Allan G., Farrell P. J., Kolman J. L. 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. doi: 10.1128/jvi.66.1.496-504.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rowe M., Rowe D. T., Gregory C. D., Young L. S., Farrell P. J., Rupani H., Rickinson A. B. Differences in B cell growth phenotype reflect novel patterns of Epstein-Barr virus latent gene expression in Burkitt's lymphoma cells. EMBO J. 1987 Sep;6(9):2743–2751. doi: 10.1002/j.1460-2075.1987.tb02568.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sakai D. D., Helms S., Carlstedt-Duke J., Gustafsson J. A., Rottman F. M., Yamamoto K. R. Hormone-mediated repression: a negative glucocorticoid response element from the bovine prolactin gene. Genes Dev. 1988 Sep;2(9):1144–1154. doi: 10.1101/gad.2.9.1144. [DOI] [PubMed] [Google Scholar]
- Sample J., Brooks L., Sample C., Young L., Rowe M., Gregory C., Rickinson A., Kieff E. Restricted Epstein-Barr virus protein expression in Burkitt lymphoma is due to a different Epstein-Barr nuclear antigen 1 transcriptional initiation site. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6343–6347. doi: 10.1073/pnas.88.14.6343. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sample J., Hummel M., Braun D., Birkenbach M., Kieff E. Nucleotide sequences of mRNAs encoding Epstein-Barr virus nuclear proteins: a probable transcriptional initiation site. Proc Natl Acad Sci U S A. 1986 Jul;83(14):5096–5100. doi: 10.1073/pnas.83.14.5096. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schaefer B. C., Strominger J. L., Speck S. H. The Epstein-Barr virus BamHI F promoter is an early lytic promoter: lack of correlation with EBNA 1 gene transcription in group 1 Burkitt's lymphoma cell lines. J Virol. 1995 Aug;69(8):5039–5047. doi: 10.1128/jvi.69.8.5039-5047.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schaefer B. C., Woisetschlaeger M., Strominger J. L., Speck S. H. Exclusive expression of Epstein-Barr virus nuclear antigen 1 in Burkitt lymphoma arises from a third promoter, distinct from the promoters used in latently infected lymphocytes. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6550–6554. doi: 10.1073/pnas.88.15.6550. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schuster C., Chasserot-Golaz S., Urier G., Beck G., Sergeant A. Evidence for a functional glucocorticoid responsive element in the Epstein-Barr virus genome. Mol Endocrinol. 1991 Feb;5(2):267–272. doi: 10.1210/mend-5-2-267. [DOI] [PubMed] [Google Scholar]
- Sinclair A. J., Brimmell M., Farrell P. J. Reciprocal antagonism of steroid hormones and BZLF1 in switch between Epstein-Barr virus latent and productive cycle gene expression. J Virol. 1992 Jan;66(1):70–77. doi: 10.1128/jvi.66.1.70-77.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sinclair A. J., Brimmell M., Shanahan F., Farrell P. J. Pathways of activation of the Epstein-Barr virus productive cycle. J Virol. 1991 May;65(5):2237–2244. doi: 10.1128/jvi.65.5.2237-2244.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sinclair A. J., Farrell P. J. Host cell requirements for efficient infection of quiescent primary B lymphocytes by Epstein-Barr virus. J Virol. 1995 Sep;69(9):5461–5468. doi: 10.1128/jvi.69.9.5461-5468.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sinclair A. J., Jacquemin M. G., Brooks L., Shanahan F., Brimmell M., Rowe M., Farrell P. J. Reduced signal transduction through glucocorticoid receptor in Burkitt's lymphoma cell lines. Virology. 1994 Mar;199(2):339–353. doi: 10.1006/viro.1994.1132. [DOI] [PubMed] [Google Scholar]
- Smith P. R., Griffin B. E. Transcription of the Epstein-Barr virus gene EBNA-1 from different promoters in nasopharyngeal carcinoma and B-lymphoblastoid cells. J Virol. 1992 Feb;66(2):706–714. doi: 10.1128/jvi.66.2.706-714.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Speck S. H., Pfitzner A., Strominger J. L. An Epstein-Barr virus transcript from a latently infected, growth-transformed B-cell line encodes a highly repetitive polypeptide. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9298–9302. doi: 10.1073/pnas.83.24.9298. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sugden B., Warren N. A promoter of Epstein-Barr virus that can function during latent infection can be transactivated by EBNA-1, a viral protein required for viral DNA replication during latent infection. J Virol. 1989 Jun;63(6):2644–2649. doi: 10.1128/jvi.63.6.2644-2649.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sung N. S., Kenney S., Gutsch D., Pagano J. S. EBNA-2 transactivates a lymphoid-specific enhancer in the BamHI C promoter of Epstein-Barr virus. J Virol. 1991 May;65(5):2164–2169. doi: 10.1128/jvi.65.5.2164-2169.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Swaminathan S., Hesselton R., Sullivan J., Kieff E. Epstein-Barr virus recombinants with specifically mutated BCRF1 genes. J Virol. 1993 Dec;67(12):7406–7413. doi: 10.1128/jvi.67.12.7406-7413.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Swaminathan S., Tomkinson B., Kieff E. Recombinant Epstein-Barr virus with small RNA (EBER) genes deleted transforms lymphocytes and replicates in vitro. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1546–1550. doi: 10.1073/pnas.88.4.1546. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tierney R. J., Steven N., Young L. S., Rickinson A. B. Epstein-Barr virus latency in blood mononuclear cells: analysis of viral gene transcription during primary infection and in the carrier state. J Virol. 1994 Nov;68(11):7374–7385. doi: 10.1128/jvi.68.11.7374-7385.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tun T., Hamaguchi Y., Matsunami N., Furukawa T., Honjo T., Kawaichi M. Recognition sequence of a highly conserved DNA binding protein RBP-J kappa. Nucleic Acids Res. 1994 Mar 25;22(6):965–971. doi: 10.1093/nar/22.6.965. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Walls D., Gannon F. The expression of novel antigens from the Epstein-Barr virus large internal repeat. EMBO J. 1988 Apr;7(4):1191–1196. doi: 10.1002/j.1460-2075.1988.tb02930.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Walls D., Perricaudet M. Novel downstream elements upregulate transcription initiated from an Epstein-Barr virus latent promoter. EMBO J. 1991 Jan;10(1):143–151. doi: 10.1002/j.1460-2075.1991.tb07930.x. [DOI] [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. doi: 10.1002/j.1460-2075.1994.tb06901.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wang F., Petti L., Braun D., Seung S., Kieff E. A bicistronic Epstein-Barr virus mRNA encodes two nuclear proteins in latently infected, growth-transformed lymphocytes. J Virol. 1987 Apr;61(4):945–954. doi: 10.1128/jvi.61.4.945-954.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Woisetschlaeger M., Jin X. W., Yandava C. N., Furmanski L. A., Strominger J. L., Speck S. H. Role for the Epstein-Barr virus nuclear antigen 2 in viral promoter switching during initial stages of infection. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3942–3946. doi: 10.1073/pnas.88.9.3942. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Woisetschlaeger M., Strominger J. L., Speck S. H. Mutually exclusive use of viral promoters in Epstein-Barr virus latently infected lymphocytes. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6498–6502. doi: 10.1073/pnas.86.17.6498. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Woisetschlaeger M., Yandava C. N., Furmanski L. A., Strominger J. L., Speck S. H. 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. doi: 10.1073/pnas.87.5.1725. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yandava C. N., Speck S. H. Characterization of the deletion and rearrangement in the BamHI C region of the X50-7 Epstein-Barr virus genome, a mutant viral strain which exhibits constitutive BamHI W promoter activity. J Virol. 1992 Sep;66(9):5646–5650. doi: 10.1128/jvi.66.9.5646-5650.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Santen V., Cheung A., Hummel M., Kieff E. RNA encoded by the IR1-U2 region of Epstein-Barr virus DNA in latently infected, growth-transformed cells. J Virol. 1983 May;46(2):424–433. doi: 10.1128/jvi.46.2.424-433.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]