Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1979 Jan;29(1):261–274. doi: 10.1128/jvi.29.1.261-274.1979

Epstein-Barr Virus-Specific RNA III. Mapping of DNA Encoding Viral RNA in Restringent Infection

Ann L Thomas Powell 1, Walter King 1, Elliott Kieff 1
PMCID: PMC353112  PMID: 219221

Abstract

Namalwa and Raji cells, originally obtained from a Burkitt tumor biopsy, grow as continuous cell lines in vitro and contain the Epstein-Barr virus (EBV)-related nuclear antigen EBNA (B. M. Reedman and G. Klein, Int. J. Cancer 11:499-520, 1973) and RNA homologous to at least 17 and 30% of the EBV genome, respectively (S. D. Hayward and E. Kieff, J. Virol. 18:518-525, 1976; T. Orellana and E. Kieff, J. Virol. 22:321-330, 1977). The polyribosomal and polyadenylated [poly(A)+] RNA fractions of Namalwa and Raji cells are enriched for a class of viral RNA homologous to 5 to 7% of EBV DNA (Hayward and Kieff, J. Virol. 18:518-525, 1976; Orellana and Kieff, J. Virol. 22:321-330, 1977). The objective of the experiments described in this communication was to determine the location within the map of the EBV genome (D. Given and E. Kieff, J. Virol. 28:524-542, 1978) of the DNA which encodes the viral RNA in the poly(A)+ and non-polyadenylated [poly(A)−] RNA fractions of Namalwa cells. Hybridization of labeled DNA homologous to Namalwa poly(A)+ or poly(A)− RNA to blots containing EcoRI, Hsu I, or Hsu I/EcoRI double-cut fragments of EBV (B95-8) or (W91) DNA indicated that these RNAs are encoded by DNA contained primarily in the Hsu I A/EcoRI A and Hsu I B/EcoRI A fragments and, to a lesser extent, in other fragments of the EBV genome. Hybridizations of Namalwa poly(A)+ and poly(A)− RNA in solution to denatured labeled EcoRI A or B fragments, Hsu I A, B, or D fragments, and Hsu I A/EcoRI A or Bam I S fragments and of Raji polyribosomal poly(A)+ RNA to the EcoRI A fragment indicated that (i) Namalwa poly(A)+ RNA is encoded primarily by 6 × 105 daltons of a 2 × 106-dalton segment of DNA, Bam I S, which is tandemly reiterated, approximately 10 times, in the Hsu I A/EcoRI A fragment and is encoded to a lesser extent by DNA in the Hsu I B, EcoRI B, and Hsu I D fragments. Raji polyribosomal poly(A)+ RNA is encoded by a similar fraction of the EcoRI A fragment as that which encodes Namalwa poly(A)+ RNA. (ii) The fraction of the Bam I S fragment homologous to Namalwa poly(A)− RNA is similar to the fraction homologous to Namalwa poly(A)+ RNA. However, Namalwa poly(A)− RNA is homologous to a larger fraction of the DNA in the Hsu I B, Hsu I D, and EcoRI B fragments.

Full text

PDF
261

Images in this article

268Image
on p.268
269Image
on p.269

Selected References

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

  1. Adams A., Bjursell G., Kaschka-Dierich C., Lindahl T. Circular Epstein-Barr virus genomes of reduced size in a human lymphoid cell line of infectious mononucleosis origin. J Virol. 1977 May;22(2):373–380. doi: 10.1128/jvi.22.2.373-380.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adams A., Lindahl T. Epstein-Barr virus genomes with properties of circular DNA molecules in carrier cells. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1477–1481. doi: 10.1073/pnas.72.4.1477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Adams A., Lindahl T., Klein G. Linear association between cellular DNA and Epstein-Barr virus DNA in a human lymphoblastoid cell line. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2888–2892. doi: 10.1073/pnas.70.10.2888. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Andersson-Anvret M., Lindahl T. Integrated viral DNA sequences in Epstein-Barr virus-converted human lymphoma lines. J Virol. 1978 Mar;25(3):710–718. doi: 10.1128/jvi.25.3.710-718.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Desrosiers R. C., Friderici K. H., Rottman F. M. Characterization of Novikoff hepatoma mRNA methylation and heterogeneity in the methylated 5' terminus. Biochemistry. 1975 Oct 7;14(20):4367–4374. doi: 10.1021/bi00691a004. [DOI] [PubMed] [Google Scholar]
  7. Dolyniuk M., Pritchett R., Kieff E. Proteins of Epstein-Barr virus. I. Analysis of the polypeptides of purified enveloped Epstein-Barr virus. J Virol. 1976 Mar;17(3):935–949. doi: 10.1128/jvi.17.3.935-949.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dubroff L. M., Nemer M. Molecular classes of heterogeneous nuclear RNA in sea urchin embryos. J Mol Biol. 1975 Jul 5;95(3):455–476. doi: 10.1016/0022-2836(75)90203-x. [DOI] [PubMed] [Google Scholar]
  9. Frenkel N., Roizman B. Ribonucleic acid synthesis in cells infected with herpes simplex virus: controls of transcription and of RNA abundance. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2654–2658. doi: 10.1073/pnas.69.9.2654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gerber P., Monroe J. H. Studies on leukocytes growing in continuous culture derived from normal human donors. J Natl Cancer Inst. 1968 Apr;40(4):855–866. [PubMed] [Google Scholar]
  11. Gerber P., Nkrumah F. K., Pritchett R., Kieff E. Comparative studies of Epstein-Barr virus strains from Ghana and the United States. Int J Cancer. 1976 Jan 15;17(1):71–81. doi: 10.1002/ijc.2910170111. [DOI] [PubMed] [Google Scholar]
  12. Gerber P., Pritchett R. F., Kieff E. D. Antigens and DNA of a chimpanzee agent related to Epstein-Barr virus. J Virol. 1976 Sep;19(3):1090–1099. doi: 10.1128/jvi.19.3.1090-1099.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gerper P., Whang-Peng J., Monroe J. H. Transformation and chromosome changes induced by Epstein-Barr virus in normal human leukocyte cultures. Proc Natl Acad Sci U S A. 1969 Jul;63(3):740–747. doi: 10.1073/pnas.63.3.740. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Given D., Kieff E. DNA of Epstein-Barr virus. IV. Linkage map of restriction enzyme fragments of the B95-8 and W91 strains of Epstein-Barr Virus. J Virol. 1978 Nov;28(2):524–542. doi: 10.1128/jvi.28.2.524-542.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gorski J., Morrison M. R., Merkel C. G., Lingrel J. B. Size heterogeneity of polyadenylate sequences in mouse globin messenger RNA. J Mol Biol. 1974 Jun 25;86(2):363–371. doi: 10.1016/0022-2836(74)90025-4. [DOI] [PubMed] [Google Scholar]
  16. Hayward S. D., Kieff E. D. Epstein-Barr virus-specific RNA. I. Analysis of viral RNA in cellular extracts and in the polyribosomal fraction of permissive and nonpermissive lymphoblastoid cell lines. J Virol. 1976 May;18(2):518–525. doi: 10.1128/jvi.18.2.518-525.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hayward S. D., Kieff E. DNA of Epstein-Barr virus. II. Comparison of the molecular weights of restriction endonuclease fragments of the DNA of Epstein-Barr virus strains and identification of end fragments of the B95-8 strain. J Virol. 1977 Aug;23(2):421–429. doi: 10.1128/jvi.23.2.421-429.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Kawai Y., Nonoyama M., Pagano J. S. Reassociation kinetics for Epstein-Barr virus DNA: nonhomology to mammalian DNA and homology of viral DNA in various diseases. J Virol. 1973 Nov;12(5):1006–1012. doi: 10.1128/jvi.12.5.1006-1012.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Klein G., Dombos L., Gothoskar B. Sensitivity of Epstein-Barr virus (EBV) producer and non-producer human lymphoblastoid cell lines to superinfection with EB-virus. Int J Cancer. 1972 Jul 15;10(1):44–57. doi: 10.1002/ijc.2910100108. [DOI] [PubMed] [Google Scholar]
  21. Klein G., Dombos L. Relationship between the sensitivity of EBV-carrying lymphoblastoid lines to superinfection and the inducibility of the resident viral genome. Int J Cancer. 1973 Mar 15;11(2):327–337. doi: 10.1002/ijc.2910110210. [DOI] [PubMed] [Google Scholar]
  22. Klein G., Pearson G., Henle G., Henle W., Diehl V., Niederman J. C. Relation between Epstein-- Barr viral and cell membrane immunofluorescence in Burkitt tumor cells. II. Comparison of cells and sera from patients with Burkitt's lymphoma and infectious mononucleosis. J Exp Med. 1968 Nov 1;128(5):1021–1030. doi: 10.1084/jem.128.5.1021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Klein G., Pearson G., Nadkarni J. S., Nadkarni J. J., Klein E., Henle G., Henle W., Clifford P. Relation between Epstein-Barr viral and cell membrane immunofluorescence of Burkitt tumor cells. I. Dependence of cell membrane immunofluorescence on presence of EB virus. J Exp Med. 1968 Nov 1;128(5):1011–1020. doi: 10.1084/jem.128.5.1011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lindahl T., Adams A., Bjursell G., Bornkamm G. W., Kaschka-Dierich C., Jehn U. Covalently closed circular duplex DNA of Epstein-Barr virus in a human lymphoid cell line. J Mol Biol. 1976 Apr 15;102(3):511–530. doi: 10.1016/0022-2836(76)90331-4. [DOI] [PubMed] [Google Scholar]
  25. Lindahl T., Klein G., Reedman B. M., Johansson B., Singh S. Relationship between Epstein-Barr virus (EBV) DNA and the EBV-determined nuclear antigen (EBNA) in Burkitt lymphoma biopsies and other lymphoproliferative malignancies. Int J Cancer. 1974 Jun 15;13(6):764–772. doi: 10.1002/ijc.2910130605. [DOI] [PubMed] [Google Scholar]
  26. Luka J., Lindahl T., Klein G. Purification of the Epstein-Barr virus-determined nuclear antigen from Epstein-Barr virus-transformed human lymphoid cell lines. J Virol. 1978 Sep;27(3):604–611. doi: 10.1128/jvi.27.3.604-611.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Luka J., Siegert W., Klein G. Solubilization of the Epstein-Barr virus-determined nuclear antigen and its characterization as a DNA-binding protein. J Virol. 1977 Apr;22(1):1–8. doi: 10.1128/jvi.22.1.1-8.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Menezes J., Jondal M., Leibold W., Dorval G. Epstein-Barr virus interactions with human lymphocyte subpopulations: virus adsorption, kinetics of expression of Epstein-Barr virus-associated nuclear antigen, and lymphocyte transformation. Infect Immun. 1976 Feb;13(2):303–310. doi: 10.1128/iai.13.2.303-310.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Menezes J., Leibold W., Klein G. Biological differences between Epstein-Barr virus (EBV) strains with regard to lymphocyte transforming ability, superinfection and antigen induction. Exp Cell Res. 1975 May;92(2):478–484. doi: 10.1016/0014-4827(75)90404-8. [DOI] [PubMed] [Google Scholar]
  30. Miller G., Coope D., Niederman J., Pagano J. Biological properties and viral surface antigens of Burkitt lymphoma- and mononucleosis- derived strains of Epstein-Barr virus released from transformed marmoset cells. J Virol. 1976 Jun;18(3):1071–1080. doi: 10.1128/jvi.18.3.1071-1080.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Miller G., Lipman M. Comparison of the yield of infectious virus from clones of human and simian lymphoblastoid lines transformed by Epstein-Barr virus. J Exp Med. 1973 Dec 1;138(6):1398–1412. doi: 10.1084/jem.138.6.1398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Miller G., Robinson J., Heston L., Lipman M. Differences between laboratory strains of Epstein-Barr virus based on immortalization, abortive infection, and interference. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4006–4010. doi: 10.1073/pnas.71.10.4006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Nilsson K. High-frequency establishment of human immunoglobulin-producing lymphoblastoid lines from normal and malignant lymphoid tissue and peripheral blood. Int J Cancer. 1971 Nov 15;8(3):432–442. doi: 10.1002/ijc.2910080311. [DOI] [PubMed] [Google Scholar]
  35. Nonoyama M., Pagano J. S. Homology between Epstein-Barr virus DNA and viral DNA from Burkitt's lymphoma and nasopharyngeal carcinoma determined by DNA-DNA reassociation kinetics. Nature. 1973 Mar 2;242(5392):44–47. doi: 10.1038/242044a0. [DOI] [PubMed] [Google Scholar]
  36. Nonoyama M., Pagano J. S. Separation of Epstein-Barr virus DNA from large chromosomal DNA in non-virus-producing cells. Nat New Biol. 1972 Aug 9;238(84):169–171. doi: 10.1038/newbio238169a0. [DOI] [PubMed] [Google Scholar]
  37. Ohno S., Luka J., Lindahl T., Klein G. Identification of a purified complement-fixing antigen as the Epstein-Barr-virus determined nuclear antigen (EBNA) by its binding to metaphase chromosomes. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1605–1609. doi: 10.1073/pnas.74.4.1605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Orellana T., Kieff E. Epstein-barr virus-specific RNA. II. Analysis of polyadenylated viral RNA in restringent, abortive, and prooductive infections. J Virol. 1977 May;22(2):321–330. doi: 10.1128/jvi.22.2.321-330.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Pope J. H. Establishment of cell lines from peripheral leucocytes in infectious mononucleosis. Nature. 1967 Nov 25;216(5117):810–811. doi: 10.1038/216810a0. [DOI] [PubMed] [Google Scholar]
  40. Pope J. H., Horne M. K., Scott W. Identification of the filtrable leukocyte-transforming factor of QIMR-WIL cells as herpes-like virus. Int J Cancer. 1969 May 15;4(3):255–260. doi: 10.1002/ijc.2910040302. [DOI] [PubMed] [Google Scholar]
  41. 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]
  42. Pritchett R. F., Hayward S. D., Kieff E. D. DNA of Epstein-Barr virus. I. Comparative studies of the DNA of Epstein-Barr virus from HR-1 and B95-8 cells: size, structure, and relatedness. J Virol. 1975 Mar;15(3):556–559. doi: 10.1128/jvi.15.3.556-559.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Pritchett R., Pendersen M., Kieff E. Complexity of EBV homologous DNA in continous lymphoblastoid cell lines. Virology. 1976 Oct 1;74(1):227–231. [PubMed] [Google Scholar]
  44. Raab-Traub N., Pritchett R., Kieff E. DNA of Epstein-Barr virus. III. Identification of restriction enzyme fragments that contain DNA sequences which differ among strains of Epstein-Barr virus. J Virol. 1978 Aug;27(2):388–398. doi: 10.1128/jvi.27.2.388-398.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Reedman B. M., Klein G. Cellular localization of an Epstein-Barr virus (EBV)-associated complement-fixing antigen in producer and non-producer lymphoblastoid cell lines. Int J Cancer. 1973 May;11(3):499–520. doi: 10.1002/ijc.2910110302. [DOI] [PubMed] [Google Scholar]
  46. Reedman B. M., Klein G., Pope J. H., Walters M. K., Hilgers J., Singh S., Johansson B. Epstein-Barr virus-associated complement-fixing and nuclear antigens in Burkitt lymphoma biopsies. Int J Cancer. 1974 Jun 15;13(6):755–763. doi: 10.1002/ijc.2910130604. [DOI] [PubMed] [Google Scholar]
  47. Rymo L., Forsblom S. Cleavage of Epstein-Barr virus DNA by restriction endonucleases EcoRI, HindIII and BamI. Nucleic Acids Res. 1978 Apr;5(4):1387–1402. doi: 10.1093/nar/5.4.1387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Seeburg P. H., Shine J., Martial J. A., Ullrich A., Baxter J. D., Goodman H. M. Nucleotide sequence of part of the gene for human chorionic somatomammotropin: purification of DNA complementary to predominant mRNA species. Cell. 1977 Sep;12(1):157–165. doi: 10.1016/0092-8674(77)90193-3. [DOI] [PubMed] [Google Scholar]
  49. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  50. WARNER R. C., SAMUELS H. H., ABBOTT M. T., KRAKOW J. S. Ribonucleic acid polymerase of Azotobacter vinelandii, II. Formation of DNA-RNA hybrids with single-stranded DNA as primer. Proc Natl Acad Sci U S A. 1963 Apr;49:533–538. doi: 10.1073/pnas.49.4.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Zimmerman S. B., Sandeen D. The ribonuclease activity of crystallized pancreatic deoxyribonuclease. Anal Biochem. 1966 Feb;14(2):269–277. doi: 10.1016/0003-2697(66)90137-0. [DOI] [PubMed] [Google Scholar]
  52. Zur Hausen H., Schulte-Holthausen H. Presence of EB virus nucleic acid homology in a "virus-free" line of Burkitt tumour cells. Nature. 1970 Jul 18;227(5255):245–248. doi: 10.1038/227245a0. [DOI] [PubMed] [Google Scholar]
  53. zur Hausen H., Schulte-Holthausen H., Klein G., Henle W., Henle G., Clifford P., Santesson L. EBV DNA in biopsies of Burkitt tumours and anaplastic carcinomas of the nasopharynx. Nature. 1970 Dec 12;228(5276):1056–1058. doi: 10.1038/2281056a0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES