Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1982 Oct;44(1):134–143. doi: 10.1128/jvi.44.1.134-143.1982

Recombination Occurs Mainly Between Parental Genomes and Precedes DNA Replication in Pseudorabies Virus-Infected Cells

Tamar Ben-Porat 1, Linda Brown 1, Ruth Ann Veach 1
PMCID: PMC256247  PMID: 6292481

Abstract

The experiments described in this paper were part of an attempt to determine the mechanisms involved in the isomerization of the pseudorabies virus genome. To this end, [14C]thymidine-labeled parental virus DNA that was transferred to progeny virions produced by cells incubated in medium containing bromodeoxy-uridine was analyzed in neutral and alkaline CsCl density gradients. The buoyant density of the 14C-labeled DNA indicated that the parental DNA strands had retained their integrity and had not undergone breakage and reunion with progeny DNA strands; neither massive intermolecular nor intramolecular recombination had occurred after replication of the DNA. Whereas breakage and reunion between parental and progeny virus DNA strands were not detectable, these processes were observed between differentially density-labeled parental DNAs. Furthermore, the frequency of recombination between progeny DNAs accumulating in the cells was low. These results indicate that in pseudorabies virus-infected rabbit kidney cells recombination occurs mainly between parental genomes and precedes DNA replication. An analysis of the kinetics of appearance of recombinants between pairwise combinations of temperature-sensitive mutants also indicated that recombination is an early event. The ratio between the number of recombinant virions and the number of temperature-sensitive mutant virions produced by the cells remained the same throughout infection. Since the relative amounts of viral DNAs synthesized early and late during the infective process that were integrated into virions were approximately the same, it appears that late viral DNA did not experience an increased number of recombinational events compared with early viral DNA. These results, which reinforce the conclusion reached from the results of the analysis of the behavior of the parental DNA molecules in density shift experiments, indicate that recombination is an early event.

Full text

PDF
134

Selected References

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

  1. BEN-PORAT T., KAPLAN A. S. The chemical composition of herpes simplex and pseudorabies viruses. Virology. 1962 Mar;16:261–266. doi: 10.1016/0042-6822(62)90246-5. [DOI] [PubMed] [Google Scholar]
  2. Becker Y. A model for intramolecular recombination of herpesvirus DNA leading to the formation of circular, circular-linear and fragmented genomes. J Theor Biol. 1978 Dec 7;75(3):339–347. doi: 10.1016/0022-5193(78)90339-9. [DOI] [PubMed] [Google Scholar]
  3. Becker Y., Asher Y., Friedmann A., Kessler E. Circular, circular-linear and branched herpes simplex virus DNA molecules from arginine deprived cells. J Gen Virol. 1978 Dec;41(3):629–633. doi: 10.1099/0022-1317-41-3-629. [DOI] [PubMed] [Google Scholar]
  4. Ben-Porat T., Blankenship M. L., DeMarchi J. M., Kaplan A. S. Replication of herpesvirus DNA. III. Rate of DNA elongation. J Virol. 1977 Jun;22(3):734–741. doi: 10.1128/jvi.22.3.734-741.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ben-Porat T., Kaplan A. S. A comparison of two populations of defective, interfering pseudorabies virus particles. Virology. 1976 Jul 15;72(2):471–479. doi: 10.1016/0042-6822(76)90175-6. [DOI] [PubMed] [Google Scholar]
  6. Ben-Porat T., Kaplan A. S. Abnormal replication of the DNA of defective pseudorabies virions. IARC Sci Publ. 1975;(11 Pt 1):97–102. [PubMed] [Google Scholar]
  7. Ben-Porat T. Replication of herpesvirus DNA. Curr Top Microbiol Immunol. 1981;91:81–107. doi: 10.1007/978-3-642-68058-8_4. [DOI] [PubMed] [Google Scholar]
  8. Ben-Porat T., Rixon F. J., Blankenship M. L. Analysis of the structure of the genome of pseudorabies virus. Virology. 1979 Jun;95(2):285–294. doi: 10.1016/0042-6822(79)90484-7. [DOI] [PubMed] [Google Scholar]
  9. Ben-Porat T., Rixon F. J. Replication of herpesvirus DNA. IV: analysis of concatemers. Virology. 1979 Apr 15;94(1):61–70. doi: 10.1016/0042-6822(79)90438-0. [DOI] [PubMed] [Google Scholar]
  10. Ben-Porat T., Stehn B., Kaplan A. S. Fate of parental herpesvirus DNA. Virology. 1976 Jun;71(2):412–422. doi: 10.1016/0042-6822(76)90369-x. [DOI] [PubMed] [Google Scholar]
  11. Ben-Porat T., Tokazewski S. A. Replication of herpesvirus DNA. II. Sedimentation characteristics of newly synthesized DNA. Virology. 1977 Jun 15;79(2):292–301. doi: 10.1016/0042-6822(77)90356-7. [DOI] [PubMed] [Google Scholar]
  12. Brown S. M., Ritchie D. A. Genetic studies with herpes simplex virus type 1. Analysis of mixed plaque-forming virus and its bearing on genetic recombination. Virology. 1975 Mar;64(1):32–42. doi: 10.1016/0042-6822(75)90076-8. [DOI] [PubMed] [Google Scholar]
  13. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  14. KAPLAN A. S., BEN-PORAT T. MODE OF REPLICATION OF PSEUDORABIES VIRUS DNA. Virology. 1964 May;23:90–95. doi: 10.1016/s0042-6822(64)80011-8. [DOI] [PubMed] [Google Scholar]
  15. KAPLAN A. S., BEN-PORAT T. The action of 5-fluorouracil on the nucleic acid metabolism of pseudorabies virus-infected and noninfected rabbit kidney cells. Virology. 1961 Jan;13:78–92. doi: 10.1016/0042-6822(61)90034-4. [DOI] [PubMed] [Google Scholar]
  16. KAPLAN A. S., VATTER A. E. A comparison of herpes simplex and pseudorabies viruses. Virology. 1959 Apr;7(4):394–407. doi: 10.1016/0042-6822(59)90068-6. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Pringle C. R., Howard D. K., Hay J. Termperature-sensitive mutants of pseudorabies virus with differential effects on viral and host DNA synthesis. Virology. 1973 Oct;55(2):495–505. doi: 10.1016/0042-6822(73)90191-8. [DOI] [PubMed] [Google Scholar]
  19. Ritchie D. A., Brown S. M., Subak-Sharpe J. H., Jamieson A. T. Heterozygosis and genetic recombination in herpes simplex type 1 virus. Virology. 1977 Oct 15;82(2):323–333. doi: 10.1016/0042-6822(77)90007-1. [DOI] [PubMed] [Google Scholar]
  20. Sheldrick P., Berthelot N. Inverted repetitions in the chromosome of herpes simplex virus. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 2):667–678. doi: 10.1101/sqb.1974.039.01.080. [DOI] [PubMed] [Google Scholar]
  21. Smiley J. R., Fong B. S., Leung W. C. Construction of a double-jointed herpes simplex viral DNA molecule: inverted repeats are required for segment inversion, and direct repeats promote deletions. Virology. 1981 Aug;113(1):345–362. doi: 10.1016/0042-6822(81)90161-6. [DOI] [PubMed] [Google Scholar]
  22. Wadsworth S., Jacob R. J., Roizman B. Anatomy of herpes simplex virus DNA. II. Size, composition, and arrangement of inverted terminal repetitions. J Virol. 1975 Jun;15(6):1487–1497. doi: 10.1128/jvi.15.6.1487-1497.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES