Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1984 May;3(5):1083–1090. doi: 10.1002/j.1460-2075.1984.tb01933.x

Transcription and DNA sequence of the BamHI L fragment of B95-8 Epstein-Barr virus.

M Biggin, P J Farrell, B G Barrell
PMCID: PMC557477  PMID: 6203743

Abstract

The sequence of the BamHI L fragment of B95-8 Epstein-Barr virus (EBV) DNA has been determined. The transcription starts of five promoters have been mapped to this fragment, using S1 mapping and either in vitro transcription or the primer extension technique. Dramatically increased levels of cytoplasmic poly(A) + RNAs, transcribed from these promoters, occur after treatment of B95-8 cells with 12-O-tetradecanoylphorbol-13-acetate (TPA). For three of these promoters addition of phosphonoacetic acid (PAA) inhibits the effect of TPA, indicating that they give rise to late lytic cycle RNAs. The other two promoters give rise to early RNAs. Northern blot analysis indicates that one of the late promoters initiates two transcripts whose size differences are due to different splicing patterns. These two RNAs code for the 350/300 and 250/200 kd envelope glycoproteins of EBV. The sequences of these proteins would be of use in the production of a synthetic vaccine to prevent EBV infection.

Full text

PDF
1083

Images in this article

Selected References

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

  1. Berk A. J., Sharp P. A. Spliced early mRNAs of simian virus 40. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1274–1278. doi: 10.1073/pnas.75.3.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bina-Stein M., Thoren M., Salzman N., Thomspon J. A. Rapid sequence determination of late simian virus 40 16S mRNA leader by using inhibitors of reverse transcriptase. Proc Natl Acad Sci U S A. 1979 Feb;76(2):731–735. doi: 10.1073/pnas.76.2.731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dambaugh T., Beisel C., Hummel M., King W., Fennewald S., Cheung A., Heller M., Raab-Traub N., Kieff E. Epstein-Barr virus (B95-8) DNA VII: molecular cloning and detailed mapping. Proc Natl Acad Sci U S A. 1980 May;77(5):2999–3003. doi: 10.1073/pnas.77.5.2999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Deininger P. L. Random subcloning of sonicated DNA: application to shotgun DNA sequence analysis. Anal Biochem. 1983 Feb 15;129(1):216–223. doi: 10.1016/0003-2697(83)90072-6. [DOI] [PubMed] [Google Scholar]
  5. Duckworth M. L., Gait M. J., Goelet P., Hong G. F., Singh M., Titmas R. C. Rapid synthesis of oligodeoxyribonucleotides VI. Efficient, mechanised synthesis of heptadecadeoxyribonucleotides by an improved solid phase phosphotriester route. Nucleic Acids Res. 1981 Apr 10;9(7):1691–1706. doi: 10.1093/nar/9.7.1691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Edson C. M., Thorley-Lawson D. A. Synthesis and processing of the three major envelope glycoproteins of Epstein-Barr virus. J Virol. 1983 May;46(2):547–556. doi: 10.1128/jvi.46.2.547-556.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. England T. E., Bruce A. G., Uhlenbeck O. C. Specific labeling of 3' termini of RNA with T4 RNA ligase. Methods Enzymol. 1980;65(1):65–74. doi: 10.1016/s0076-6879(80)65011-3. [DOI] [PubMed] [Google Scholar]
  8. Farrell P. J., Deininger P. L., Bankier A., Barrell B. Homologous upstream sequences near Epstein-Barr virus promoters. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1565–1569. doi: 10.1073/pnas.80.6.1565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hoffman G. J., Lazarowitz S. G., Hayward S. D. Monoclonal antibody against a 250,000-dalton glycoprotein of Epstein-Barr virus identifies a membrane antigen and a neutralizing antigen. Proc Natl Acad Sci U S A. 1980 May;77(5):2979–2983. doi: 10.1073/pnas.77.5.2979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hummel M., Kieff E. Epstein-Barr virus RNA. VIII. Viral RNA in permissively infected B95-8 cells. J Virol. 1982 Jul;43(1):262–272. doi: 10.1128/jvi.43.1.262-272.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Hummel M., Thorley-Lawson D., Kieff E. An Epstein-Barr virus DNA fragment encodes messages for the two major envelope glycoproteins (gp350/300 and gp220/200). J Virol. 1984 Feb;49(2):413–417. doi: 10.1128/jvi.49.2.413-417.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kozak M. Possible role of flanking nucleotides in recognition of the AUG initiator codon by eukaryotic ribosomes. Nucleic Acids Res. 1981 Oct 24;9(20):5233–5252. doi: 10.1093/nar/9.20.5233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Manley J. L., Fire A., Cano A., Sharp P. A., Gefter M. L. DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3855–3859. doi: 10.1073/pnas.77.7.3855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  17. Miller G., Shope T., Lisco H., Stitt D., Lipman M. Epstein-Barr virus: transformation, cytopathic changes, and viral antigens in squirrel monkey and marmoset leukocytes. Proc Natl Acad Sci U S A. 1972 Feb;69(2):383–387. doi: 10.1073/pnas.69.2.383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Morgan A. J., North J. R., Epstein M. A. Purification and properties of the gp340 component of Epstein-Barr virus membrane antigen in an immunogenic form. J Gen Virol. 1983 Feb;64(Pt 2):455–460. doi: 10.1099/0022-1317-64-2-455. [DOI] [PubMed] [Google Scholar]
  19. North J. R., Morgan A. J., Thompson J. L., Epstein M. A. Purified Epstein-Barr virus Mr 340,000 glycoprotein induces potent virus-neutralizing antibodies when incorporated in liposomes. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7504–7508. doi: 10.1073/pnas.79.23.7504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  21. Sanger F., Coulson A. R., Barrell B. G., Smith A. J., Roe B. A. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. doi: 10.1016/0022-2836(80)90196-5. [DOI] [PubMed] [Google Scholar]
  22. Sanger F., Coulson A. R. The use of thin acrylamide gels for DNA sequencing. FEBS Lett. 1978 Mar 1;87(1):107–110. doi: 10.1016/0014-5793(78)80145-8. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Skare J., Strominger J. L. Cloning and mapping of BamHi endonuclease fragments of DNA from the transforming B95-8 strain of Epstein-Barr virus. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3860–3864. doi: 10.1073/pnas.77.7.3860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Staden R. Automation of the computer handling of gel reading data produced by the shotgun method of DNA sequencing. Nucleic Acids Res. 1982 Aug 11;10(15):4731–4751. doi: 10.1093/nar/10.15.4731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Staden R. Graphic methods to determine the function of nucleic acid sequences. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):521–538. doi: 10.1093/nar/12.1part2.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Thorley-Lawson D. A., Geilinger K. Monoclonal antibodies against the major glycoprotein (gp350/220) of Epstein-Barr virus neutralize infectivity. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5307–5311. doi: 10.1073/pnas.77.9.5307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Thorley-Lawson D. A., Poodry C. A. Identification and isolation of the main component (gp350-gp220) of Epstein-Barr virus responsible for generating neutralizing antibodies in vivo. J Virol. 1982 Aug;43(2):730–736. doi: 10.1128/jvi.43.2.730-736.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Weigel R., Miller G. Major EB virus-specific cytoplasmic transcripts in a cellular clone of the HR-1 Burkitt lymphoma line during latency and after induction of viral replicative cycle by phorbol esters. Virology. 1983 Mar;125(2):287–298. doi: 10.1016/0042-6822(83)90202-7. [DOI] [PubMed] [Google Scholar]
  31. Weil P. A., Luse D. S., Segall J., Roeder R. G. Selective and accurate initiation of transcription at the Ad2 major late promotor in a soluble system dependent on purified RNA polymerase II and DNA. Cell. 1979 Oct;18(2):469–484. doi: 10.1016/0092-8674(79)90065-5. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES