Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1997 Jun;71(6):4209–4217. doi: 10.1128/jvi.71.6.4209-4217.1997

Circularization and cleavage of guinea pig cytomegalovirus genomes.

M A McVoy 1, D E Nixon 1, S P Adler 1
PMCID: PMC191635  PMID: 9151807

Abstract

The mechanisms by which herpesvirus genome ends are fused to form circles after infection and are re-formed by cleavage from concatemeric DNA are unknown. We used the simple structure of guinea pig cytomegalovirus genomes, which have either one repeated DNA sequence at each end or one repeat at one end and no repeat at the other, to study these mechanisms. In circular DNA, two restriction fragments contained fused terminal sequences and had sizes consistent with the presence of single or double terminal repeats. This result implies a simple ligation of genomic ends and shows that circularization does not occur by annealing of single-stranded terminal repeats formed by exonuclease digestion. Cleavage to form the two genome types occurred at two sites, and homologies between these sites identified two potential cis elements that may be necessary for cleavage. One element coincided with the A-rich region of a pac2 sequence and had 9 of 11 bases identical between the two sites. The second element had six bases identical at both sites, in each case 7 bp from the termini. To confirm the presence of cis cleavage elements, a recombinant virus in which foreign sequences displaced the 6- and 11-bp elements 1 kb from the cleavage point was constructed. Cleavage at the disrupted site did not occur. In a second recombinant virus, restoration of 64 bases containing the 6- and 11-bp elements to the disrupted cleavage site restored cleavage. Therefore, cis cleavage elements exist within this 64-base region, and sequence conservation suggests that they are the 6- and 11-bp elements.

Full Text

The Full Text of this article is available as a PDF (918.9 KB).

Selected References

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

  1. Barnett J. W., Eppstein D. A., Chan H. W. Class I defective herpes simplex virus DNA as a molecular cloning vehicle in eucaryotic cells. J Virol. 1983 Nov;48(2):384–395. doi: 10.1128/jvi.48.2.384-395.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chowdhury S. I., Buhk H. J., Ludwig H., Hammerschmidt W. Genomic termini of equine herpesvirus 1. J Virol. 1990 Feb;64(2):873–880. doi: 10.1128/jvi.64.2.873-880.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Davison A. J. Structure of the genome termini of varicella-zoster virus. J Gen Virol. 1984 Nov;65(Pt 11):1969–1977. doi: 10.1099/0022-1317-65-11-1969. [DOI] [PubMed] [Google Scholar]
  4. Deiss L. P., Chou J., Frenkel N. Functional domains within the a sequence involved in the cleavage-packaging of herpes simplex virus DNA. J Virol. 1986 Sep;59(3):605–618. doi: 10.1128/jvi.59.3.605-618.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Deiss L. P., Frenkel N. Herpes simplex virus amplicon: cleavage of concatemeric DNA is linked to packaging and involves amplification of the terminally reiterated a sequence. J Virol. 1986 Mar;57(3):933–941. doi: 10.1128/jvi.57.3.933-941.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gao M., Isom H. C. Characterization of the guinea pig cytomegalovirus genome by molecular cloning and physical mapping. J Virol. 1984 Nov;52(2):436–447. doi: 10.1128/jvi.52.2.436-447.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Garber D. A., Beverley S. M., Coen D. M. Demonstration of circularization of herpes simplex virus DNA following infection using pulsed field gel electrophoresis. Virology. 1993 Nov;197(1):459–462. doi: 10.1006/viro.1993.1612. [DOI] [PubMed] [Google Scholar]
  8. Grafstrom R. H., Alwine J. C., Steinhart W. L., Hill C. W., Hyman R. W. The terminal repetition of herpes simplex virus DNA. Virology. 1975 Sep;67(1):144–157. doi: 10.1016/0042-6822(75)90412-2. [DOI] [PubMed] [Google Scholar]
  9. Hammerschmidt W., Ludwig H., Buhk H. J. Specificity of cleavage in replicative-form DNA of bovine herpesvirus 1. J Virol. 1988 Apr;62(4):1355–1363. doi: 10.1128/jvi.62.4.1355-1363.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Harper L., Demarchi J., Ben-Porat T. Sequence of the genome ends and of the junction between the ends in concatemeric DNA of pseudorabies virus. J Virol. 1986 Dec;60(3):1183–1185. doi: 10.1128/jvi.60.3.1183-1185.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jean J. H., Ben-Porat T. Appearance in vivo of single-stranded complementary ends on parental herpesvirus DNA. Proc Natl Acad Sci U S A. 1976 Aug;73(8):2674–2678. doi: 10.1073/pnas.73.8.2674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kudler L., Hyman R. W. Exonuclease III digestion of herpes simplex virus DNA. Virology. 1979 Jan 15;92(1):68–81. doi: 10.1016/0042-6822(79)90215-0. [DOI] [PubMed] [Google Scholar]
  13. Marks J. R., Spector D. H. Replication of the murine cytomegalovirus genome: structure and role of the termini in the generation and cleavage of concatenates. Virology. 1988 Jan;162(1):98–107. doi: 10.1016/0042-6822(88)90398-4. [DOI] [PubMed] [Google Scholar]
  14. McVoy M. A., Adler S. P. Human cytomegalovirus DNA replicates after early circularization by concatemer formation, and inversion occurs within the concatemer. J Virol. 1994 Feb;68(2):1040–1051. doi: 10.1128/jvi.68.2.1040-1051.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mocarski E. S., Liu A. C., Spaete R. R. Structure and variability of the a sequence in the genome of human cytomegalovirus (Towne strain). J Gen Virol. 1987 Aug;68(Pt 8):2223–2230. doi: 10.1099/0022-1317-68-8-2223. [DOI] [PubMed] [Google Scholar]
  16. Mocarski E. S., Post L. E., Roizman B. Molecular engineering of the herpes simplex virus genome: insertion of a second L-S junction into the genome causes additional genome inversions. Cell. 1980 Nov;22(1 Pt 1):243–255. doi: 10.1016/0092-8674(80)90172-5. [DOI] [PubMed] [Google Scholar]
  17. Mocarski E. S., Roizman B. Site-specific inversion sequence of the herpes simplex virus genome: domain and structural features. Proc Natl Acad Sci U S A. 1981 Nov;78(11):7047–7051. doi: 10.1073/pnas.78.11.7047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mocarski E. S., Roizman B. Structure and role of the herpes simplex virus DNA termini in inversion, circularization and generation of virion DNA. Cell. 1982 Nov;31(1):89–97. doi: 10.1016/0092-8674(82)90408-1. [DOI] [PubMed] [Google Scholar]
  19. Nasseri M., Mocarski E. S. The cleavage recognition signal is contained within sequences surrounding an a-a junction in herpes simplex virus DNA. Virology. 1988 Nov;167(1):25–30. doi: 10.1016/0042-6822(88)90050-5. [DOI] [PubMed] [Google Scholar]
  20. Secchiero P., Nicholas J., Deng H., Xiaopeng T., van Loon N., Ruvolo V. R., Berneman Z. N., Reitz M. S., Jr, Dewhurst S. Identification of human telomeric repeat motifs at the genome termini of human herpesvirus 7: structural analysis and heterogeneity. J Virol. 1995 Dec;69(12):8041–8045. doi: 10.1128/jvi.69.12.8041-8045.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Smiley J. R., Duncan J., Howes M. Sequence requirements for DNA rearrangements induced by the terminal repeat of herpes simplex virus type 1 KOS DNA. J Virol. 1990 Oct;64(10):5036–5050. doi: 10.1128/jvi.64.10.5036-5050.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Spaete R. R., Frenkel N. The herpes simplex virus amplicon: a new eucaryotic defective-virus cloning-amplifying vector. Cell. 1982 Aug;30(1):295–304. doi: 10.1016/0092-8674(82)90035-6. [DOI] [PubMed] [Google Scholar]
  23. Stow N. D., McMonagle E. C., Davison A. J. Fragments from both termini of the herpes simplex virus type 1 genome contain signals required for the encapsidation of viral DNA. Nucleic Acids Res. 1983 Dec 10;11(23):8205–8220. doi: 10.1093/nar/11.23.8205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tamashiro J. C., Filpula D., Friedmann T., Spector D. H. Structure of the heterogeneous L-S junction region of human cytomegalovirus strain AD169 DNA. J Virol. 1984 Nov;52(2):541–548. doi: 10.1128/jvi.52.2.541-548.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tamashiro J. C., Spector D. H. Terminal structure and heterogeneity in human cytomegalovirus strain AD169. J Virol. 1986 Sep;59(3):591–604. doi: 10.1128/jvi.59.3.591-604.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tenser R. B., Hsiung G. D. Comparison of guinea pig cytomegalovirus and guinea pig herpes-like virus: pathogenesis and persistence in experimentally infected animals. Infect Immun. 1976 Mar;13(3):934–940. doi: 10.1128/iai.13.3.934-940.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Thomson B. J., Dewhurst S., Gray D. Structure and heterogeneity of the a sequences of human herpesvirus 6 strain variants U1102 and Z29 and identification of human telomeric repeat sequences at the genomic termini. J Virol. 1994 May;68(5):3007–3014. doi: 10.1128/jvi.68.5.3007-3014.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Varmuza S. L., Smiley J. R. Signals for site-specific cleavage of HSV DNA: maturation involves two separate cleavage events at sites distal to the recognition sequences. Cell. 1985 Jul;41(3):793–802. doi: 10.1016/s0092-8674(85)80060-x. [DOI] [PubMed] [Google Scholar]
  29. Vink C., Beuken E., Bruggeman C. A. Structure of the rat cytomegalovirus genome termini. J Virol. 1996 Aug;70(8):5221–5229. doi: 10.1128/jvi.70.8.5221-5229.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wadsworth S., Hayward G. S., Roizman B. Anatomy of herpes simplex virus DNA. V. Terminally repetitive sequences. J Virol. 1976 Feb;17(2):503–512. doi: 10.1128/jvi.17.2.503-512.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Zimmermann J., Hammerschmidt W. Structure and role of the terminal repeats of Epstein-Barr virus in processing and packaging of virion DNA. J Virol. 1995 May;69(5):3147–3155. doi: 10.1128/jvi.69.5.3147-3155.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES