Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1979 Jan;29(1):83–90. doi: 10.1128/jvi.29.1.83-90.1979

Herpes Simplex Virus Type 2 Functions Expressed During Stimulation of Human Cell DNA Synthesis

Louis S Kucera 1, Iris Edwards 1
PMCID: PMC353078  PMID: 219236

Abstract

Experiments were designed to identify herpes simplex virus type 2 (HSV-2)-specific functions expressed during stimulation of human embryo fibroblast DNA synthesis. Cultures were partially arrested in DNA synthesis by pretreatment with 5-fluorouracil and maintenance in low-serum (0.2%) medium during virus infection. Results showed that continuous [methyl-3H]thymidine uptake into cellular DNA was ninefold greater in HSV-2-infected than in mock-infected cultures measured after 24 h of incubation at 42°C. Shifting mock-infected cultures from low- to high-serum (10%) medium also caused some stimulation, but [methyl-3H]thymidine uptake was only twofold greater than in cells maintained with low serum. Plating efficiencies of both HSV-2-infected and mock-infected cells at 42°C were essentially the same and ranged from 37 to 76% between zero time and 72 h of incubation. De novo RNA and protein syntheses were continuously required for HSV-2 stimulation of cellular DNA synthesis. HSV-2 infection markedly enhanced transport, phosphorylation, and rate of incorporation of [methyl-3H]thymidine into cellular DNA, starting at 3 h and reaching a maximum by 12 h; after 12 h, these processes gradually declined to low levels. In mock-infected cells these processes remained at low levels throughout the observation period. Pretreatment of cells with interferon or addition of arabinofuranosylthymine at the time of virus infection inhibited stimulation caused by HSV-2. 5-Bromodeoxyuridine density-labeled experiments revealed that HSV-2 stimulates predominantly semiconservative DNA replication and some DNA repair. Stimulation of [methyl-3H]thymidine into cellular DNA correlated with detection of virus-specific thymidine kinase activity. In conclusion, HSV-2 stimulation of cellular DNA synthesis appeared to involve at least four virus-specific functions: induction of thymidine transport, HSV-2 thymidine kinase activity, semiconservative replication, and repair of cellular DNA.

Full text

PDF
83

Selected References

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

  1. Aswell J. F., Gentry G. A. Cell-dependent antiherpesviral activity of 5-methylarabinosylcytosine, an intracellular ara-T donor. Ann N Y Acad Sci. 1977 Mar 4;284:342–350. doi: 10.1111/j.1749-6632.1977.tb21969.x. [DOI] [PubMed] [Google Scholar]
  2. Bittlingmaier K., Schneider D., Falke D. Thymidine transport in herpesvirus hominis type 1 and 2 infected BHK 21 cells. J Gen Virol. 1977 Apr;35(1):159–173. doi: 10.1099/0022-1317-35-1-159. [DOI] [PubMed] [Google Scholar]
  3. Breslow R. E., Goldsby R. A. Isolation and characterization of thymidine transport mutants of Chinese hamster cells. Exp Cell Res. 1969 Jun;55(3):339–346. doi: 10.1016/0014-4827(69)90567-9. [DOI] [PubMed] [Google Scholar]
  4. DULBECCO R., HARTWELL L. H., VOGT M. INDUCTION OF CELLULAR DNA SYNTHESIS BY POLYOMA VIRUS. Proc Natl Acad Sci U S A. 1965 Feb;53:403–410. doi: 10.1073/pnas.53.2.403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fried M., Pitts J. D. Replication of polyoma virus DNA. I. A resting cell system for biochemical studies on polyoma virus. Virology. 1968 Apr;34(4):761–770. doi: 10.1016/0042-6822(68)90097-4. [DOI] [PubMed] [Google Scholar]
  6. Gershon D., Sachs L., Winocour E. The induction of cellular DNA synthesis by simian virus 40 in contact-inhibited and in x-irradiated cells. Proc Natl Acad Sci U S A. 1966 Sep;56(3):918–925. doi: 10.1073/pnas.56.3.918. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Glasgow L. A., Hanshaw J. B., Merigan T. C., Petralli J. K. Interferon and cytomegalovirus in vivo and in vitro. Proc Soc Exp Biol Med. 1967 Jul;125(3):843–849. doi: 10.3181/00379727-125-32220. [DOI] [PubMed] [Google Scholar]
  8. Hatanaka M., Dulbecco R. SV 40-specific thymidine kinase. Proc Natl Acad Sci U S A. 1967 Nov;58(5):1888–1894. doi: 10.1073/pnas.58.5.1888. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Henry P., Black P. H., Oxman M. N., Weissman S. M. Stimulation of DNA synthesis in mouse cell line 3T3 by Simian virus 40. Proc Natl Acad Sci U S A. 1966 Oct;56(4):1170–1176. doi: 10.1073/pnas.56.4.1170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Kucera L. S. Effects of temperature on frog polyhedral cytoplasmic deoxyribovirus multiplication: thermosensitivity of initiation, replication, and encapsidation of viral DNA. Virology. 1970 Nov;42(3):576–589. doi: 10.1016/0042-6822(70)90304-1. [DOI] [PubMed] [Google Scholar]
  12. Kucera L. S., Gusdon J. P. Transformation of human embryonic fibroblasts by photodynamically inactivated herpes simplex virus, type 2 at supra-optimal temperature. J Gen Virol. 1976 Feb;30(2):257–261. doi: 10.1099/0022-1317-30-2-257. [DOI] [PubMed] [Google Scholar]
  13. Larsson A., Reichard P. Enzymatic synthesis of deoxyribonucleotides. X. Reduction of purine ribonucleotides; allosteric behavior and substrate specificity of the enzyme system from Escherichia coli B. J Biol Chem. 1966 Jun 10;241(11):2540–2549. [PubMed] [Google Scholar]
  14. Lindberg U., Nordenskjöld B. A., Reichard P., Skoog L. Thymidine phosphate pools and DNA synthesis after polyoma infection of mouse embryo cells. Cancer Res. 1969 Aug;29(8):1498–1506. [PubMed] [Google Scholar]
  15. Lorentz A. K., Munk K., Darai G. DNA repair replication in human embryonic lung cells infected with herpes simplex virus. Virology. 1977 Oct 15;82(2):401–408. doi: 10.1016/0042-6822(77)90015-0. [DOI] [PubMed] [Google Scholar]
  16. Marcon M. J., Kucera L. S. Consequences of herpes simplex virus type 2 and human cell interaction at supraoptimal temperatures. J Virol. 1976 Oct;20(1):54–62. doi: 10.1128/jvi.20.1.54-62.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Melvin P., Kucera L. S. Induction of human cell DNA synthesis by herpes simplex virus type 2. J Virol. 1975 Mar;15(3):534–539. doi: 10.1128/jvi.15.3.534-539.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Miller R. L., Iltis J. P., Rapp F. Differential effect of arabinofuranosylthymine of the replication of human herpesviruses. J Virol. 1977 Sep;23(3):679–684. doi: 10.1128/jvi.23.3.679-684.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Plagemann P. G., Richey D. P. Transport of nucleosides, nucleic acid bases, choline and glucose by animal cells in culture. Biochim Biophys Acta. 1974 Dec 16;344(3-4):263–305. doi: 10.1016/0304-4157(74)90010-0. [DOI] [PubMed] [Google Scholar]
  20. ROIZMAN B., ROANE P. R., Jr THE MULTIPLICATION OF HERPES SIMPLEX VIRUS. II. THE RELATION BETWEEN PROTEIN SYNTHESIS AND THE DUPLICATION OF VIRAL DNA IN INFECTED HEP-2 CELLS. Virology. 1964 Feb;22:262–269. doi: 10.1016/0042-6822(64)90011-x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES