Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1976 Nov;20(2):413–424. doi: 10.1128/jvi.20.2.413-424.1976

Herpes simplex virus gene expression in transformed cells. I. Regulation of the viral thymidine kinase gene in transformed L cells by products of superinfecting virus.

J M Leiden, R Buttyan, P G Spear
PMCID: PMC355009  PMID: 185425

Abstract

In this paper we show that the expression of the herpes simplex virus type 1 (HSV-1) gene for thymidine kinase (tk) in HSV-transformed cells is subject to regulation by two viral products synthesized during productive infection of these cells with a tk- mutant of HSV-1. The cell line used in this study is a derivative of tk-deficient mouse L cells that, after exposure to UV-inactivated HSV-1, had acquired the HSV-1 gene for tk (which we term a resident viral gene) and consequently expressed the tk+ phenotype (LVtk+ cells). Productive infection of these cells with HSV-1(tk-) at appropriate multiplicities caused significant enhancement of the viral tk activity. The results of several experiments allow us to conclude that this enhancement was due to increased synthesis of tk specified by the HSV-1 gene resident in the LVtk+ cells and that a specific protein made early after infection with HSV-1(tk-) mediated the enhancement, probably by increasing the production of mRNA from the viral tk gene resident in the LVtk+ cells. Our data also indicate that another HSV-1(tk-) product acted to turn off tk synthesis. The finding that tk activity continued to increase for a longer time after infection of the LVtk+ cells at 2 PFU/cell than after infection at higher multiplicities suggested the synthesis of a product which inhibited tk synthesis and whose concentration reached critical levels earlier at higher multiplicities of infection. Inhibition of DNA synthesis after infection, a treatment that depresses the synthesis of late viral proteins, prolonged the synthesis of tk in LVtk+ cells infected at either 2 or 5 PFU/cell. Infection of the LVtk+ cells with HSV-2(tk-) resulted in only small increases in tk activity, indicating some type specificity in recognition of viral products that can modify the expression of the HSV-1 tk gene resident in these cells.

Full text

PDF
413

Selected References

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

  1. Chadha K. C., Munyon W. Presence of herpes simplex virus-related antigens in transformed L cells. J Virol. 1975 Jun;15(6):1475–1486. doi: 10.1128/jvi.15.6.1475-1486.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Copple C. D., McDougall J. K. Clonal derivatives of a herpes type 2 transformed hamster cell line (333-8-9): cytogenetic analysis, tumorigenicity and virus sequence detection. Int J Cancer. 1976 Apr 15;17(4):501–510. doi: 10.1002/ijc.2910170413. [DOI] [PubMed] [Google Scholar]
  3. DUBBS D. R., KIT S. MUTANT STRAINS OF HERPES SIMPLEX DEFICIENT IN THYMIDINE KINASE-INDUCING ACTIVITY. Virology. 1964 Apr;22:493–502. doi: 10.1016/0042-6822(64)90070-4. [DOI] [PubMed] [Google Scholar]
  4. Davidson R. L., Adelstein S. J., Oxman M. N. Herpes simplex virus as a source of thymidine kinase for thymidine kinase-deficient mouse cells: suppression and reactivation of the viral enzyme. Proc Natl Acad Sci U S A. 1973 Jul;70(7):1912–1916. doi: 10.1073/pnas.70.7.1912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Davis D. B., Munyon W., Buchsbaum R., Chawda R. Virus type-specific thymidine kinase in cells biochemically transformed by herpes simplex virus types 1 and 2. J Virol. 1974 Jan;13(1):140–145. doi: 10.1128/jvi.13.1.140-145.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Duff R., Rapp F. Oncogenic transformation of hamster embryo cells after exposure to inactivated herpes simplex virus type 1. J Virol. 1973 Aug;12(2):209–217. doi: 10.1128/jvi.12.2.209-217.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Duff R., Rapp F. Properties of hamster embryo fibroblasts transformed in vitro after exposure to ultraviolet-irradiated herpes simplex virus type 2. J Virol. 1971 Oct;8(4):469–477. doi: 10.1128/jvi.8.4.469-477.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Frenkel N., Locker H., Cox B., Roizman B., Rapp F. Herpes simplex virus DNA in transformed cells: sequence complexity in five hamster cell lines and one derived hamster tumor. J Virol. 1976 Jun;18(3):885–893. doi: 10.1128/jvi.18.3.885-893.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Garfinkle B., McAuslan B. R. Regulation of herpes simplex virus-induced thymidine kinase. Biochem Biophys Res Commun. 1974 Jun 4;58(3):822–829. doi: 10.1016/s0006-291x(74)80491-2. [DOI] [PubMed] [Google Scholar]
  10. Honess R. W., Roizman B. Proteins specified by herpes simplex virus. XI. Identification and relative molar rates of synthesis of structural and nonstructural herpes virus polypeptides in the infected cell. J Virol. 1973 Dec;12(6):1347–1365. doi: 10.1128/jvi.12.6.1347-1365.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Honess R. W., Roizman B. Regulation of herpesvirus macromolecular synthesis. I. Cascade regulation of the synthesis of three groups of viral proteins. J Virol. 1974 Jul;14(1):8–19. doi: 10.1128/jvi.14.1.8-19.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Honess R. W., Roizman B. Regulation of herpesvirus macromolecular synthesis: sequential transition of polypeptide synthesis requires functional viral polypeptides. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1276–1280. doi: 10.1073/pnas.72.4.1276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Honess R. W., Watson D. H. Absence of a requirement for host polypeptides in the herpes virus thymidine kinase. J Gen Virol. 1974 Jul;24(1):215–220. doi: 10.1099/0022-1317-24-1-215. [DOI] [PubMed] [Google Scholar]
  14. Honess R. W., Watson D. H. Herpes simplex virus-specific polypeptides studied by polyacrylamide gel electrophoresis of immune precipitates. J Gen Virol. 1974 Feb;22(2):171–185. doi: 10.1099/0022-1317-22-2-171. [DOI] [PubMed] [Google Scholar]
  15. Huang E. S. Human cytomegalovirus. IV. Specific inhibition of virus-induced DNA polymerase activity and viral DNA replication by phosphonoacetic acid. J Virol. 1975 Dec;16(6):1560–1565. doi: 10.1128/jvi.16.6.1560-1565.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jamieson A. T., Subak-Sharpe J. H. Biochemical studies on the herpes simplex virus-specified deoxypyrimidine kinase activity. J Gen Virol. 1974 Sep;24(3):481–492. doi: 10.1099/0022-1317-24-3-481. [DOI] [PubMed] [Google Scholar]
  17. KIT S., DUBBS D. R., PIEKARSKI L. J., HSU T. C. DELETION OF THYMIDINE KINASE ACTIVITY FROM L CELLS RESISTANT TO BROMODEOXYURIDINE. Exp Cell Res. 1963 Aug;31:297–312. doi: 10.1016/0014-4827(63)90007-7. [DOI] [PubMed] [Google Scholar]
  18. Kit S., Dubbs D. R., Anken M. Altered properties of thymidine kinase after infection of mouse fibroblast cells with herpes simplex virus. J Virol. 1967 Feb;1(1):238–240. doi: 10.1128/jvi.1.1.238-240.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kit S., Leung W. C., Jorgensen G. N., Dubbs D. R. Distinctive properties of thymidine kinase isozymes induced by human and avian hepresviruses. Int J Cancer. 1974 Nov 15;14(5):598–610. doi: 10.1002/ijc.2910140506. [DOI] [PubMed] [Google Scholar]
  20. Kit S., Leung W. C., Trkula D., Jorgensen G. Gel electrophoresis and isoelectric focusing of mitochondrial and viral-induced thymidine kinases. Int J Cancer. 1974 Feb 15;13(2):203–218. doi: 10.1002/ijc.2910130208. [DOI] [PubMed] [Google Scholar]
  21. Kozak M., Roizman B. Regulation of herpesvirus macromolecular synthesis: nuclear retention of nontranslated viral RNA sequences. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4322–4326. doi: 10.1073/pnas.71.11.4322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kraiselburd E., Gage L. P., Weissbach A. Presence of a herpes simplex virus DNA fragment in an L cell clone obtained after infection with irradiated herpes simplex virus I. J Mol Biol. 1975 Oct 5;97(4):533–542. doi: 10.1016/s0022-2836(75)80057-x. [DOI] [PubMed] [Google Scholar]
  23. LITTLEFIELD J. W. SELECTION OF HYBRIDS FROM MATINGS OF FIBROBLASTS IN VITRO AND THEIR PRESUMED RECOMBINANTS. Science. 1964 Aug 14;145(3633):709–710. doi: 10.1126/science.145.3633.709. [DOI] [PubMed] [Google Scholar]
  24. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  25. Leinbach S. S., Reno J. M., Lee L. F., Isbell A. F., Boezi J. A. Mechanism of phosphonoacetate inhibition of herpesvirus-induced DNA polymerase. Biochemistry. 1976 Jan 27;15(2):426–430. doi: 10.1021/bi00647a029. [DOI] [PubMed] [Google Scholar]
  26. Lin S. S., Munyon W. Expression of the viral thymidine kinase gene in herpes simplex virus-transformed L cells. J Virol. 1974 Nov;14(5):1199–1208. doi: 10.1128/jvi.14.5.1199-1208.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Macnab J. C. Transformation of rat embryo cells by temperature-sensitive mutants of herpes simplex virus. J Gen Virol. 1974 Jul;24(1):143–153. doi: 10.1099/0022-1317-24-1-143. [DOI] [PubMed] [Google Scholar]
  28. Mao J. C., Robishaw E. E., Overby L. R. Inhibition of DNA polymerase from herpes simplex virus-infected wi-38 cells by phosphonoacetic Acid. J Virol. 1975 May;15(5):1281–1283. doi: 10.1128/jvi.15.5.1281-1283.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Minson A. C., Thouless M. E., Eglin R. P., Darby G. The detection of virus DNA sequences in a herpes type 2 transformed hamster cell line (333-8-9). Int J Cancer. 1976 Apr 15;17(4):493–500. doi: 10.1002/ijc.2910170412. [DOI] [PubMed] [Google Scholar]
  30. Munyon W., Buchsbaum R., Paoletti E., Mann J., Kraiselburd E., Davis D. Electrophoresis of thymidine kinase activity synthesized by cells transformed by herpes simplex virus. Virology. 1972 Sep;49(3):683–689. doi: 10.1016/0042-6822(72)90525-9. [DOI] [PubMed] [Google Scholar]
  31. Munyon W., Kraiselburd E., Davis D., Mann J. Transfer of thymidine kinase to thymidine kinaseless L cells by infection with ultraviolet-irradiated herpes simplex virus. J Virol. 1971 Jun;7(6):813–820. doi: 10.1128/jvi.7.6.813-820.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Márquez E. D., Rapp F. Presence of herpesvirus-associated antigens on the surfaces of transformed, tumor, and metastatic cells and enhanced antigenicity of transformed cells using 5-iodo-2'-deoxyuridine. Intervirology. 1975;6(2):64–71. doi: 10.1159/000149457. [DOI] [PubMed] [Google Scholar]
  33. Powell K. L., Courtney R. J. Polypeptide synthesized in herpes simplex virus type 2-infected HEp-2 cells. Virology. 1975 Jul;66(1):217–228. doi: 10.1016/0042-6822(75)90192-0. [DOI] [PubMed] [Google Scholar]
  34. ROIZMAN B., ROANE P. R., Jr A physical difference between two strains of herpes simplex virus apparent on sedimentation in cesium chloride. Virology. 1961 Sep;15:75–79. doi: 10.1016/0042-6822(61)90079-4. [DOI] [PubMed] [Google Scholar]
  35. Rapp F., Buss E. R. Comparison of herpes simplex virus isolates using a quantitative selection assay for transformation. Intervirology. 1975;6(2):72–82. doi: 10.1159/000149458. [DOI] [PubMed] [Google Scholar]
  36. Reed C. L., Cohen G. H., Rapp F. Detection of a virus-specific antigen on the surface of herpes simplex virus-transformed cells. J Virol. 1975 Mar;15(3):668–670. doi: 10.1128/jvi.15.3.668-670.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Roizman B., Kozak M., Honess R. W., Hayward G. Regulation of herpesvirus macromolecular synthesis: evidence for multilevel regulation of herpes simplex 1 RNA and protein synthesis. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 2):687–701. doi: 10.1101/sqb.1974.039.01.083. [DOI] [PubMed] [Google Scholar]
  38. Roizman B., Spear P. G. Preparation of herpes simplex virus of high titer. J Virol. 1968 Jan;2(1):83–84. doi: 10.1128/jvi.2.1.83-84.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Spear P. G., Roizman B. The proteins specified by herpes simplex virus. I. Time of synthesis, transfer into nuclei, and properties of proteins made in productively infected cells. Virology. 1968 Dec;36(4):545–555. doi: 10.1016/0042-6822(68)90186-4. [DOI] [PubMed] [Google Scholar]
  40. Summers W. P., Wagner M., Summers W. C. Possible peptide chain termination mutants in thymide kinase gene of a mammalian virus, herpes simplex virus. Proc Natl Acad Sci U S A. 1975 Oct;72(10):4081–4084. doi: 10.1073/pnas.72.10.4081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Thouless M. E., Chadha K. C., Munyon W. H. Serological specificity of thymidine kinase activity in herpes simplex virus-transformed L cells. Virology. 1976 Jan;69(1):350–351. doi: 10.1016/0042-6822(76)90225-7. [DOI] [PubMed] [Google Scholar]
  42. Thouless M. E. Serological properties of thymidine kinase produced in cells infected with type 1 or type 2 herpes virus. J Gen Virol. 1972 Dec;17(3):307–315. doi: 10.1099/0022-1317-17-3-307. [DOI] [PubMed] [Google Scholar]
  43. Thouless M. E., Wildy P. Deoxypyrimidine kinases of herpes simplex viruses types 1 and 2: comparison of serological and structural properties. J Gen Virol. 1975 Feb;26(2):159–170. doi: 10.1099/0022-1317-26-2-159. [DOI] [PubMed] [Google Scholar]

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

RESOURCES