Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1989 Nov;63(11):4609–4615. doi: 10.1128/jvi.63.11.4609-4615.1989

Identification and characterization of an Epstein-Barr virus early antigen that is encoded by the NotI repeats.

C M Nuebling 1, N Mueller-Lantzsch 1
PMCID: PMC251094  PMID: 2552144

Abstract

The Epstein-Barr virus (EBV) genome is characterized by two regions carrying partially homologous clusters of short tandem repeats (NotI and PstI repeats) flanked by 1,044 and 1,045 base pairs with almost perfect homology (DL and DR, left and right duplications, respectively). Both repetitive regions are transcribed into poly(A)+ mRNA after induction of the productive EBV cycle with the tumor promoter 12-O-tetradecanoylphorbol-13-acetate and contain open reading frames. To identify the potential protein encoded by the NotI repeat open reading frame (BHLF1), two repeat units of EBV strain M-ABA were expressed using the tryptophan-regulated Escherichia coli expression vector pATH11. Rabbit antisera generated against the resulting fusion protein reacted specifically with a protein varying in molecular size between 70,000 and 90,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, found after 12-O-tetradecanoyl-phorbol-13-acetate or n-butyrate induction in various cell lines harboring EBV. In immunofluorescence tests with the BHLF1-specific antiserum, an immunofluorescence with EA-D specificity could be observed. In addition, the BHLF1 protein is exhibiting polyanion-binding activity with a maximum for single-stranded DNA. Furthermore, the fusion protein is recognized by a number of human EBV-positive sera.

Full text

PDF
4609

Images in this article

4611Image
on p.4611
4611Image
on p.4611
4612Image
on p.4612
4612Image
on p.4612
4612Image
on p.4612
4613Image
on p.4613

Selected References

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

  1. 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]
  2. Bornkamm G. W., Delius H., Zimber U., Hudewentz J., Epstein M. A. Comparison of Epstein-Barr virus strains of different origin by analysis of the viral DNAs. J Virol. 1980 Sep;35(3):603–618. doi: 10.1128/jvi.35.3.603-618.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bornkamm G. W., Hudewentz J., Freese U. K., Zimber U. Deletion of the nontransforming Epstein-Barr virus strain P3HR-1 causes fusion of the large internal repeat to the DSL region. J Virol. 1982 Sep;43(3):952–968. doi: 10.1128/jvi.43.3.952-968.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Crawford D. H., Epstein M. A., Bornkamm G. W., Achong B. G., Finerty S., Thompson J. L. Biological and biochemical observations on isolates of EB virus from the malignant epithelial cells of two nasopharyngeal carcinomas. Int J Cancer. 1979 Sep 15;24(3):294–302. doi: 10.1002/ijc.2910240305. [DOI] [PubMed] [Google Scholar]
  5. Dambaugh T. R., Kieff E. Identification and nucleotide sequences of two similar tandem direct repeats in Epstein-Barr virus DNA. J Virol. 1982 Dec;44(3):823–833. doi: 10.1128/jvi.44.3.823-833.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. DePamphilis M. L. Transcriptional elements as components of eukaryotic origins of DNA replication. Cell. 1988 Mar 11;52(5):635–638. doi: 10.1016/0092-8674(88)90398-4. [DOI] [PubMed] [Google Scholar]
  7. Freese U. K., Laux G., Hudewentz J., Schwarz E., Bornkamm G. W. Two distant clusters of partially homologous small repeats of Epstein-Barr virus are transcribed upon induction of an abortive or lytic cycle of the virus. J Virol. 1983 Dec;48(3):731–743. doi: 10.1128/jvi.48.3.731-743.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fresen K. O., Hausen H. Establishment of EBNA-expressing cell lines by infection of Epstein-Barr virus (EBV)-genome-negative human lymphoma cells with different EBV strains. Int J Cancer. 1976 Feb 15;17(2):161–166. doi: 10.1002/ijc.2910170203. [DOI] [PubMed] [Google Scholar]
  9. Gergely L., Klein G., Ernberg I. The action of DNA antagonists on Epstein-Barr virus (EBV)-associated early antigen (EA) in Burkitt lymphoma lines. Int J Cancer. 1971 Mar 15;7(2):293–302. doi: 10.1002/ijc.2910070214. [DOI] [PubMed] [Google Scholar]
  10. Hammerschmidt W., Sugden B. Identification and characterization of oriLyt, a lytic origin of DNA replication of Epstein-Barr virus. Cell. 1988 Nov 4;55(3):427–433. doi: 10.1016/0092-8674(88)90028-1. [DOI] [PubMed] [Google Scholar]
  11. Hayward S. D., Lazarowitz S. G., Hayward G. S. Organization of the Epstein-Barr virus DNA molecule. II. Fine mapping of the boundaries of the internal repeat cluster of B95-8 and identification of additional small tandem repeats adjacent to the HR-1 deletion. J Virol. 1982 Jul;43(1):201–212. doi: 10.1128/jvi.43.1.201-212.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Henle W., Diehl V., Kohn G., Zur Hausen H., Henle G. Herpes-type virus and chromosome marker in normal leukocytes after growth with irradiated Burkitt cells. Science. 1967 Sep 1;157(3792):1064–1065. doi: 10.1126/science.157.3792.1064. [DOI] [PubMed] [Google Scholar]
  13. Hudewentz J., Delius H., Freese U. K., Zimber U., Bornkamm G. W. Two distant regions of the Epstein-Barr virus genome with sequence homologies have the same orientation and involve small tandem repeats. EMBO J. 1982;1(1):21–26. doi: 10.1002/j.1460-2075.1982.tb01118.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hummel M., Kieff E. Mapping of polypeptides encoded by the Epstein-Barr virus genome in productive infection. Proc Natl Acad Sci U S A. 1982 Sep;79(18):5698–5702. doi: 10.1073/pnas.79.18.5698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jeang K. T., Hayward S. D. Organization of the Epstein-Barr virus DNA molecule. III. Location of the P3HR-1 deletion junction and characterization of the NotI repeat units that form part of the template for an abundant 12-O-tetradecanoylphorbol-13-acetate-induced mRNA transcript. J Virol. 1983 Oct;48(1):135–148. doi: 10.1128/jvi.48.1.135-148.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. Laux G., Freese U. K., Bornkamm G. W. Structure and evolution of two related transcription units of Epstein-Barr virus carrying small tandem repeats. J Virol. 1985 Dec;56(3):987–995. doi: 10.1128/jvi.56.3.987-995.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Leinbach S. S., Reno J. M., Lee L. F., Isbell A. F., Boezi J. A. Mechanism of phosphonoacetate inhibition of herpesvirus-induced DNA polymerase. Biochemistry. 1976 Jan 27;15(2):426–430. doi: 10.1021/bi00647a029. [DOI] [PubMed] [Google Scholar]
  20. Lieberman P. M., Hardwick J. M., Hayward S. D. Responsiveness of the Epstein-Barr virus NotI repeat promoter to the Z transactivator is mediated in a cell-type-specific manner by two independent signal regions. J Virol. 1989 Jul;63(7):3040–3050. doi: 10.1128/jvi.63.7.3040-3050.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Luka J., Kallin B., Klein G. Induction of the Epstein-Barr virus (EBV) cycle in latently infected cells by n-butyrate. Virology. 1979 Apr 15;94(1):228–231. doi: 10.1016/0042-6822(79)90455-0. [DOI] [PubMed] [Google Scholar]
  22. Maurizi M. R., Trisler P., Gottesman S. Insertional mutagenesis of the lon gene in Escherichia coli: lon is dispensable. J Bacteriol. 1985 Dec;164(3):1124–1135. doi: 10.1128/jb.164.3.1124-1135.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pfitzner A. J., Tsai E. C., Strominger J. L., Speck S. H. Isolation and characterization of cDNA clones corresponding to transcripts from the BamHI H and F regions of the Epstein-Barr virus genome. J Virol. 1987 Sep;61(9):2902–2909. doi: 10.1128/jvi.61.9.2902-2909.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Polack A., Hartl G., Zimber U., Freese U. K., Laux G., Takaki K., Hohn B., Gissmann L., Bornkamm G. W. A complete set of overlapping cosmid clones of M-ABA virus derived from nasopharyngeal carcinoma and its similarity to other Epstein-Barr virus isolates. Gene. 1984 Mar;27(3):279–288. doi: 10.1016/0378-1119(84)90072-6. [DOI] [PubMed] [Google Scholar]
  25. Pope J. H., Horne M. K., Scott W. Transformation of foetal human keukocytes in vitro by filtrates of a human leukaemic cell line containing herpes-like virus. Int J Cancer. 1968 Nov 15;3(6):857–866. doi: 10.1002/ijc.2910030619. [DOI] [PubMed] [Google Scholar]
  26. Raab-Traub N., Dambaugh T., Kieff E. DNA of Epstein-Barr virus VIII: B95-8, the previous prototype, is an unusual deletion derivative. Cell. 1980 Nov;22(1 Pt 1):257–267. doi: 10.1016/0092-8674(80)90173-7. [DOI] [PubMed] [Google Scholar]
  27. Roeckel D., Mueller-Lantzsch N. Biochemical characterization of two Epstein-Barr virus early antigen-associated phosphopolypeptides. Virology. 1985 Dec;147(2):253–263. doi: 10.1016/0042-6822(85)90128-x. [DOI] [PubMed] [Google Scholar]
  28. Schauder B., Blöcker H., Frank R., McCarthy J. E. Inducible expression vectors incorporating the Escherichia coli atpE translational initiation region. Gene. 1987;52(2-3):279–283. doi: 10.1016/0378-1119(87)90054-0. [DOI] [PubMed] [Google Scholar]
  29. Scheidtmann K. H., Hardung M., Echle B., Walter G. DNA-binding activity of simian virus 40 large T antigen correlates with a distinct phosphorylation state. J Virol. 1984 Apr;50(1):1–12. doi: 10.1128/jvi.50.1.1-12.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Seibl R., Wolf H. Mapping of Epstein-Barr virus proteins on the genome by translation of hybrid-selected RNA from induced P3HR1 cells and induced Raji cells. Virology. 1985 Feb;141(1):1–13. doi: 10.1016/0042-6822(85)90177-1. [DOI] [PubMed] [Google Scholar]
  31. Spindler K. R., Rosser D. S., Berk A. J. Analysis of adenovirus transforming proteins from early regions 1A and 1B with antisera to inducible fusion antigens produced in Escherichia coli. J Virol. 1984 Jan;49(1):132–141. doi: 10.1128/jvi.49.1.132-141.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Walter G., Scheidtmann K. H., Carbone A., Laudano A. P., Doolittle R. F. Antibodies specific for the carboxy- and amino-terminal regions of simian virus 40 large tumor antigen. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5197–5200. doi: 10.1073/pnas.77.9.5197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. White R. A., Wilcox M. Protein products of the bithorax complex in Drosophila. Cell. 1984 Nov;39(1):163–171. doi: 10.1016/0092-8674(84)90202-2. [DOI] [PubMed] [Google Scholar]
  34. Yamamoto M., Tabata T., Smith M., Tanaka A., Nonoyama M. Cycloheximide-resistant gene of Epstein-Barr virus in freshly infected B lymphocytes. Virology. 1989 May;170(1):307–310. doi: 10.1016/0042-6822(89)90385-1. [DOI] [PubMed] [Google Scholar]
  35. Zimber U., Adldinger H. K., Lenoir G. M., Vuillaume M., Knebel-Doeberitz M. V., Laux G., Desgranges C., Wittmann P., Freese U. K., Schneider U. Geographical prevalence of two types of Epstein-Barr virus. Virology. 1986 Oct 15;154(1):56–66. doi: 10.1016/0042-6822(86)90429-0. [DOI] [PubMed] [Google Scholar]
  36. zur Hausen H., Bornkamm G. W., Schmidt R., Hecker E. Tumor initiators and promoters in the induction of Epstein-Barr virus. Proc Natl Acad Sci U S A. 1979 Feb;76(2):782–785. doi: 10.1073/pnas.76.2.782. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. zur Hausen H., O'Neill F. J., Freese U. K., Hecker E. Persisting oncogenic herpesvirus induced by the tumour promotor TPA. Nature. 1978 Mar 23;272(5651):373–375. doi: 10.1038/272373a0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES