Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1996 Jun;70(6):3423–3431. doi: 10.1128/jvi.70.6.3423-3431.1996

Plasmid-like replicative intermediates of the Epstein-Barr virus lytic origin of DNA replication.

R Pfüller 1, W Hammerschmidt 1
PMCID: PMC190215  PMID: 8648674

Abstract

During the lytic phase of herpesviruses, intermediates of viral DNA replication are found as large concatemeric molecules in the infected cells. It is not known, however, what the early events in viral DNA replication that yield these concatemers are. In an attempt to identify these early steps of DNA replication, replicative intermediates derived from the lytic origin of Epstein-Barr virus, oriLyt, were analyzed. As shown by density shift experiments with bromodeoxyuridine, oriLyt replicated semiconservatively soon after induction of the lytic cycle and oriLyt-containing DNA is amplified to yield monomeric plasmid progeny DNA (besides multimeric forms and high-molecular-weight DNA). A new class of plasmid progeny DNA which have far fewer negative supercoils than do plasmids extracted from uninduced cells is present only in cells undergoing the lytic cycle of Epstein-Barr virus. This finding is consistent with plasmid DNAs having fewer nucleosomes before extraction. The newly replicated plasmid DNAs are dependent on a functional oriLyt in cis and support an efficient marker transfer into Escherichia coli as monomeric plasmids. Multimeric forms of presumably circular progeny DNA of oriLyt, as well as detected recombination events, indicate that oriLyt-mediated DNA replication is biphasic: an early theta-like mode is followed by a complex pattern which could result from rolling-circle DNA replication.

Full Text

The Full Text of this article is available as a PDF (627.0 KB).

Selected References

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

  1. Adams A. Replication of latent Epstein-Barr virus genomes in Raji cells. J Virol. 1987 May;61(5):1743–1746. doi: 10.1128/jvi.61.5.1743-1746.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Bastia D., Sueoka N. Studies on the late replication of phage lambda: rolling-circle replication of the wild type and a partially suppressed strain, Oam29 Pam80. J Mol Biol. 1975 Oct 25;98(2):305–320. doi: 10.1016/s0022-2836(75)80120-3. [DOI] [PubMed] [Google Scholar]
  4. Bataille D., Epstein A. Herpes simplex virus replicative concatemers contain L components in inverted orientation. Virology. 1994 Sep;203(2):384–388. doi: 10.1006/viro.1994.1498. [DOI] [PubMed] [Google Scholar]
  5. Carter B. J., Shaw B. D., Smith M. G. Two stages in the replication of bacteriophage lambda DNA. Biochim Biophys Acta. 1969 Dec 16;195(2):494–505. doi: 10.1016/0005-2787(69)90656-x. [DOI] [PubMed] [Google Scholar]
  6. Challberg M. D. A method for identifying the viral genes required for herpesvirus DNA replication. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9094–9098. doi: 10.1073/pnas.83.23.9094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Confalonieri F., Elie C., Nadal M., de La Tour C., Forterre P., Duguet M. Reverse gyrase: a helicase-like domain and a type I topoisomerase in the same polypeptide. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4753–4757. doi: 10.1073/pnas.90.10.4753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Countryman J., Miller G. Activation of expression of latent Epstein-Barr herpesvirus after gene transfer with a small cloned subfragment of heterogeneous viral DNA. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4085–4089. doi: 10.1073/pnas.82.12.4085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. DePamphilis M. L. Eukaryotic DNA replication: anatomy of an origin. Annu Rev Biochem. 1993;62:29–63. doi: 10.1146/annurev.bi.62.070193.000333. [DOI] [PubMed] [Google Scholar]
  10. Ebert S. N., Subramanian D., Shtrom S. S., Chung I. K., Parris D. S., Muller M. T. Association between the p170 form of human topoisomerase II and progeny viral DNA in cells infected with herpes simplex virus type 1. J Virol. 1994 Feb;68(2):1010–1020. doi: 10.1128/jvi.68.2.1010-1020.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Farrell P. J., Rowe D. T., Rooney C. M., Kouzarides T. Epstein-Barr virus BZLF1 trans-activator specifically binds to a consensus AP-1 site and is related to c-fos. EMBO J. 1989 Jan;8(1):127–132. doi: 10.1002/j.1460-2075.1989.tb03356.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fixman E. D., Hayward G. S., Hayward S. D. Replication of Epstein-Barr virus oriLyt: lack of a dedicated virally encoded origin-binding protein and dependence on Zta in cotransfection assays. J Virol. 1995 May;69(5):2998–3006. doi: 10.1128/jvi.69.5.2998-3006.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fixman E. D., Hayward G. S., Hayward S. D. trans-acting requirements for replication of Epstein-Barr virus ori-Lyt. J Virol. 1992 Aug;66(8):5030–5039. doi: 10.1128/jvi.66.8.5030-5039.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gahn T. A., Schildkraut C. L. The Epstein-Barr virus origin of plasmid replication, oriP, contains both the initiation and termination sites of DNA replication. Cell. 1989 Aug 11;58(3):527–535. doi: 10.1016/0092-8674(89)90433-9. [DOI] [PubMed] [Google Scholar]
  15. Glaser R., Nonoyama M. Host cell regulation of induction of Epstein-Barr virus. J Virol. 1974 Jul;14(1):174–176. doi: 10.1128/jvi.14.1.174-176.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gray H. B., Jr, Upholt W. B., Vinograd J. A buoyant method for the determination of the superhelix density of closed circular DNA. J Mol Biol. 1971 Nov 28;62(1):1–19. doi: 10.1016/0022-2836(71)90127-6. [DOI] [PubMed] [Google Scholar]
  17. Gruffat H., Renner O., Pich D., Hammerschmidt W. Cellular proteins bind to the downstream component of the lytic origin of DNA replication of Epstein-Barr virus. J Virol. 1995 Mar;69(3):1878–1886. doi: 10.1128/jvi.69.3.1878-1886.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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. doi: 10.1038/340393a0. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  21. Jacob R. J., Morse L. S., Roizman B. Anatomy of herpes simplex virus DNA. XII. Accumulation of head-to-tail concatemers in nuclei of infected cells and their role in the generation of the four isomeric arrangements of viral DNA. J Virol. 1979 Feb;29(2):448–457. doi: 10.1128/jvi.29.2.448-457.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jacob R. J., Roizman B. Anatomy of herpes simplex virus DNA VIII. Properties of the replicating DNA. J Virol. 1977 Aug;23(2):394–411. doi: 10.1128/jvi.23.2.394-411.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Keller W. Determination of the number of superhelical turns in simian virus 40 DNA by gel electrophoresis. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4876–4880. doi: 10.1073/pnas.72.12.4876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kikuchi A., Asai K. Reverse gyrase--a topoisomerase which introduces positive superhelical turns into DNA. Nature. 1984 Jun 21;309(5970):677–681. doi: 10.1038/309677a0. [DOI] [PubMed] [Google Scholar]
  25. Koo H. S., Lau K., Wu H. Y., Liu L. F. Identification of a DNA supercoiling activity in Saccharomyces cerevisiae. Nucleic Acids Res. 1992 Oct 11;20(19):5067–5072. doi: 10.1093/nar/20.19.5067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lee M. S., Garrard W. T. Positive DNA supercoiling generates a chromatin conformation characteristic of highly active genes. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9675–9679. doi: 10.1073/pnas.88.21.9675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lieberman P. M., Hardwick J. M., Sample J., Hayward G. S., Hayward S. D. The zta transactivator involved in induction of lytic cycle gene expression in Epstein-Barr virus-infected lymphocytes binds to both AP-1 and ZRE sites in target promoter and enhancer regions. J Virol. 1990 Mar;64(3):1143–1155. doi: 10.1128/jvi.64.3.1143-1155.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Meselson M., Stahl F. W. THE REPLICATION OF DNA IN ESCHERICHIA COLI. Proc Natl Acad Sci U S A. 1958 Jul 15;44(7):671–682. doi: 10.1073/pnas.44.7.671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Meuth M., Green H. Induction of a deoxycytidineless state in cultured mammalian cells by bromodeoxyuridine. Cell. 1974 Jun;2(2):109–112. doi: 10.1016/0092-8674(74)90099-3. [DOI] [PubMed] [Google Scholar]
  30. Pruss G. J., Drlica K. Topoisomerase I mutants: the gene on pBR322 that encodes resistance to tetracycline affects plasmid DNA supercoiling. Proc Natl Acad Sci U S A. 1986 Dec;83(23):8952–8956. doi: 10.1073/pnas.83.23.8952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Reisman D., Yates J., Sugden B. A putative origin of replication of plasmids derived from Epstein-Barr virus is composed of two cis-acting components. Mol Cell Biol. 1985 Aug;5(8):1822–1832. doi: 10.1128/mcb.5.8.1822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Roizman B. The structure and isomerization of herpes simplex virus genomes. Cell. 1979 Mar;16(3):481–494. doi: 10.1016/0092-8674(79)90023-0. [DOI] [PubMed] [Google Scholar]
  33. Schepers A., Pich D., Hammerschmidt W. A transcription factor with homology to the AP-1 family links RNA transcription and DNA replication in the lytic cycle of Epstein-Barr virus. EMBO J. 1993 Oct;12(10):3921–3929. doi: 10.1002/j.1460-2075.1993.tb06070.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schepers A., Pich D., Mankertz J., Hammerschmidt W. cis-acting elements in the lytic origin of DNA replication of Epstein-Barr virus. J Virol. 1993 Jul;67(7):4237–4245. doi: 10.1128/jvi.67.7.4237-4245.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Severini A., Morgan A. R., Tovell D. R., Tyrrell D. L. Study of the structure of replicative intermediates of HSV-1 DNA by pulsed-field gel electrophoresis. Virology. 1994 May 1;200(2):428–435. doi: 10.1006/viro.1994.1206. [DOI] [PubMed] [Google Scholar]
  36. Skaliter R., Lehman I. R. Rolling circle DNA replication in vitro by a complex of herpes simplex virus type 1-encoded enzymes. Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10665–10669. doi: 10.1073/pnas.91.22.10665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sugden B., Marsh K., Yates J. A vector that replicates as a plasmid and can be efficiently selected in B-lymphoblasts transformed by Epstein-Barr virus. Mol Cell Biol. 1985 Feb;5(2):410–413. doi: 10.1128/mcb.5.2.410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Takada K., Horinouchi K., Ono Y., Aya T., Osato T., Takahashi M., Hayasaka S. An Epstein-Barr virus-producer line Akata: establishment of the cell line and analysis of viral DNA. Virus Genes. 1991 Apr;5(2):147–156. doi: 10.1007/BF00571929. [DOI] [PubMed] [Google Scholar]
  39. Takada K., Shimizu N., Sakuma S., Ono Y. trans activation of the latent Epstein-Barr virus (EBV) genome after transfection of the EBV DNA fragment. J Virol. 1986 Mar;57(3):1016–1022. doi: 10.1128/jvi.57.3.1016-1022.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wu C. A., Nelson N. J., McGeoch D. J., Challberg M. D. Identification of herpes simplex virus type 1 genes required for origin-dependent DNA synthesis. J Virol. 1988 Feb;62(2):435–443. doi: 10.1128/jvi.62.2.435-443.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Yates J. L., Guan N. Epstein-Barr virus-derived plasmids replicate only once per cell cycle and are not amplified after entry into cells. J Virol. 1991 Jan;65(1):483–488. doi: 10.1128/jvi.65.1.483-488.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Yates J. L., Warren N., Sugden B. Stable replication of plasmids derived from Epstein-Barr virus in various mammalian cells. 1985 Feb 28-Mar 6Nature. 313(6005):812–815. doi: 10.1038/313812a0. [DOI] [PubMed] [Google Scholar]
  43. Zhang X., Efstathiou S., Simmons A. Identification of novel herpes simplex virus replicative intermediates by field inversion gel electrophoresis: implications for viral DNA amplification strategies. Virology. 1994 Aug 1;202(2):530–539. doi: 10.1006/viro.1994.1375. [DOI] [PubMed] [Google Scholar]

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

RESOURCES