Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1980 Dec;36(3):746–755. doi: 10.1128/jvi.36.3.746-755.1980

Properties of Cells Carrying the Herpes Simplex Virus Type 2 Thymidine Kinase Gene: Mechanisms of Reversion to a Thymidine Kinase-Negative Phenotype

K F Bastow 1, G Darby 1, P Wildy 1, A C Minson 1
PMCID: PMC353702  PMID: 16789205

Abstract

We have isolated cells with a thymidine kinase-negative (tk−) phenotype from cells which carry the herpes simplex virus type 2 tk gene by selection in 5-bromodeoxyuridine or 9-(2-hydroxyethoxymethyl)guanine. Both selection routines generated revertants with a frequency of 10−3 to 10−4, and resistance to either compound conferred simultaneous resistance to the other. tk− revertants fell into three classes: (i) cells that arose by deletion of all virus sequences, (ii) cells that had lost the virus tk gene but retained a nonselected virus-specific function and arose by deletion of part of the virus-specific sequence, and (iii) cells that retained the potential to express all of the virus-specific functions of the parental cells and retained all of the virus-specific DNA sequences.

Full text

PDF
753

Images in this article

750Image
on p.750
750Image
on p.750
750Image
on p.750
751Image
on p.751

Selected References

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

  1. Bacchetti S., Graham F. L. Characterization of human TK- cell lines transformed to a TK+ phenotype by herpes simplex virus type 2 DNA. J Gen Virol. 1979 Jan;42(1):149–157. doi: 10.1099/0022-1317-42-1-149. [DOI] [PubMed] [Google Scholar]
  2. Bacchetti S., Graham F. L. Transfer of the gene for thymidine kinase to thymidine kinase-deficient human cells by purified herpes simplex viral DNA. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1590–1594. doi: 10.1073/pnas.74.4.1590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buchan A., Watson D. H., Dubbs D. R., Kit S. Serological study of a mutant of herpesvirus unable to stimulate thymidine kinase. J Virol. 1970 Jun;5(6):817–818. doi: 10.1128/jvi.5.6.817-818.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chadha K. C., Munyon W. H., Hughes R. G., Jr Thymidine kinaseless revertants of Ltk- cells transformed by herpes simplex virus type 1 are resistant to retransformation by homologous virus. Infect Immun. 1977 May;16(2):655–661. doi: 10.1128/iai.16.2.655-661.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clements J. B., Watson R. J., Wilkie N. M. Temporal regulation of herpes simplex virus type 1 transcription: location of transcripts on the viral genome. Cell. 1977 Sep;12(1):275–285. doi: 10.1016/0092-8674(77)90205-7. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Deaven L. L., Petersen D. F. The chromosomes of CHO, an aneuploid Chinese hamster cell line: G-band, C-band, and autoradiographic analyses. Chromosoma. 1973;41(2):129–144. doi: 10.1007/BF00319690. [DOI] [PubMed] [Google Scholar]
  8. Fyfe J. A., Keller P. M., Furman P. A., Miller R. L., Elion G. B. Thymidine kinase from herpes simplex virus phosphorylates the new antiviral compound, 9-(2-hydroxyethoxymethyl)guanine. J Biol Chem. 1978 Dec 25;253(24):8721–8727. [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. Hicks J., Strathern J. N., Klar A. J. Transposable mating type genes in Saccharomyces cerevisiae. Nature. 1979 Nov 29;282(5738):478–473. doi: 10.1038/282478a0. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Jones P. C., Hayward G. S., Roizman B. Anatomy of herpes simplex virus DNA VII. alpha-RNA is homologous to noncontiguous sites in both the L and S components of viral DNA. J Virol. 1977 Jan;21(1):268–276. doi: 10.1128/jvi.21.1.268-276.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kadouri A., Kunce J. J., Lark K. G. Evidence for dominant mutations reducing HGPRT activity. Nature. 1978 Jul 20;274(5668):256–259. doi: 10.1038/274256a0. [DOI] [PubMed] [Google Scholar]
  14. Kaufman E. R., Davidson R. L. Control of the expression of a herpes simplex virus thymidine kinase gene incorporated into thymidine kinase-deficient mouse cells. Somatic Cell Genet. 1975 Apr;1(2):153–163. doi: 10.1007/BF01538545. [DOI] [PubMed] [Google Scholar]
  15. Kit S., Dubbs D. R. Regulation of herpesvirus thymidine kinase activity in LM(TK) cells transformed by ultraviolet light-irradiated herpes simplex virus. Virology. 1977 Jan;76(1):331–340. doi: 10.1016/0042-6822(77)90306-3. [DOI] [PubMed] [Google Scholar]
  16. Kit S., Dubbs D. R., Schaffer P. A. Thymidine kinase activity of biochemically transformed mouse cells after superinfection by thymidine kinase-negative, temperature-sensitive, herpes simplex virus mutants. Virology. 1978 Apr;85(2):456–463. doi: 10.1016/0042-6822(78)90452-x. [DOI] [PubMed] [Google Scholar]
  17. Klemperer H. G., Haynes G. R., Shedden W. I., Watson D. H. A virus-specific thymidine kinase in BHK-21 cells infected with herpes simplex virus. Virology. 1967 Jan;31(1):120–128. doi: 10.1016/0042-6822(67)90015-3. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. LITTLEFIELD J. W. STUDIES ON THYMIDINE KINASE IN CULTURED MOUSE FIBROBLASTS. Biochim Biophys Acta. 1965 Jan 11;95:14–22. doi: 10.1016/0005-2787(65)90206-6. [DOI] [PubMed] [Google Scholar]
  20. Lai E. C., Woo S. L., Bordelon-Riser M. E., Fraser T. H., O'Malley B. W. Ovalbumin is synthesized in mouse cells transformed with the natural chicken ovalbumin gene. Proc Natl Acad Sci U S A. 1980 Jan;77(1):244–248. doi: 10.1073/pnas.77.1.244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Leiden J. M., Buttyan R., Spear P. G. 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 Virol. 1976 Nov;20(2):413–424. doi: 10.1128/jvi.20.2.413-424.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Leiden J. M., Frenkel N., Rapp F. Identification of the herpes simplex virus DNA sequences present in six herpes simplex virus thymidine kinase-transformed mouse cell lines. J Virol. 1980 Jan;33(1):272–285. doi: 10.1128/jvi.33.1.272-285.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Maitland N. J., McDougall J. K. Biochemical transformation of mouse cells by fragments of herpes simplex virus DNA. Cell. 1977 May;11(1):233–241. doi: 10.1016/0092-8674(77)90334-8. [DOI] [PubMed] [Google Scholar]
  25. Mantei N., Boll W., Weissmann C. Rabbit beta-globin mRNA production in mouse L cells transformed with cloned rabbit beta-globin chromosomal DNA. Nature. 1979 Sep 6;281(5726):40–46. doi: 10.1038/281040a0. [DOI] [PubMed] [Google Scholar]
  26. Minson A. C., Darby G. K., Wildy P. Virus-specific DNA sequences present in cells which carry the herpes simplex virus thymidine kinase gene. J Gen Virol. 1979 Nov;45(2):489–496. doi: 10.1099/0022-1317-45-2-489. [DOI] [PubMed] [Google Scholar]
  27. Minson A. C., Wildy P., Buchan A., Darby G. Introduction of the herpes simplex virus thymidine kinase gene into mouse cells using virus DNA or transformed cell DNA. Cell. 1978 Mar;13(3):581–587. doi: 10.1016/0092-8674(78)90331-8. [DOI] [PubMed] [Google Scholar]
  28. Morse L. S., Pereira L., Roizman B., Schaffer P. A. Anatomy of herpes simplex virus (HSV) DNA. X. Mapping of viral genes by analysis of polypeptides and functions specified by HSV-1 X HSV-2 recombinants. J Virol. 1978 May;26(2):389–410. doi: 10.1128/jvi.26.2.389-410.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Nishiyama Y., Rapp F. Anticellular effects of 9-(2-hydroxyethoxymethyl) guanine against herpes simplex virus-transformed cells. J Gen Virol. 1979 Oct;45(1):227–230. doi: 10.1099/0022-1317-45-1-227. [DOI] [PubMed] [Google Scholar]
  31. Preston V. G., Davison A. J., Marsden H. S., Timbury M. C., Subak-Sharpe J. H., Wilkie N. M. Recombinants between herpes simplex virus types 1 and 2: analyses of genome structures and expression of immediate early polypeptides. J Virol. 1978 Nov;28(2):499–517. doi: 10.1128/jvi.28.2.499-517.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rapp F., Turner N., Schaffer P. A. Biochemical transformation by temperature-sensitive mutants of herpes simplex virus type 1. J Virol. 1980 Jun;34(3):704–710. doi: 10.1128/jvi.34.3.704-710.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Reyes G. R., LaFemina R., Hayward S. D., Hayward G. S. Morphological transformation by DNA fragments of human herpesviruses: evidence for two distinct transforming regions in herpes simplex virus types 1 and 2 and lack of correlation with biochemical transfer of the thymidine kinase gene. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 1):629–641. doi: 10.1101/sqb.1980.044.01.066. [DOI] [PubMed] [Google Scholar]
  34. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  35. Schaffer P. A., Carter V. C., Timbury M. C. Collaborative complementation study of temperature-sensitive mutants of herpes simplex virus types 1 and 2. J Virol. 1978 Sep;27(3):490–504. doi: 10.1128/jvi.27.3.490-504.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sharp J. D., Capecchi N. E., Capecchi M. R. Altered enzymes in drug-resistant variants of mammalian tissue culture cells. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3145–3149. doi: 10.1073/pnas.70.11.3145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  38. Steinberg B., Pollack R., Topp W., Botchan M. Isolation and characterization of T antigen-negative revertants from a line of transformed rat cells containing one copy of the SV40 genome. Cell. 1978 Jan;13(1):19–32. doi: 10.1016/0092-8674(78)90134-4. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Sutter D., Doerfler W. Methylation of integrated adenovirus type 12 DNA sequences in transformed cells is inversely correlated with viral gene expression. Proc Natl Acad Sci U S A. 1980 Jan;77(1):253–256. doi: 10.1073/pnas.77.1.253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Varmus H. E., Vogt P. K., Bishop J. M. Integration of deoxyribonucleic acid specific for Rous sarcoma virus after infection of permissive and nonpermissive hosts. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3067–3071. doi: 10.1073/pnas.70.11.3067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Watson R. J., Clements J. B. A herpes simplex virus type 1 function continuously required for early and late virus RNA synthesis. Nature. 1980 May 29;285(5763):329–330. doi: 10.1038/285329a0. [DOI] [PubMed] [Google Scholar]
  43. Watson R. J., Clements J. B. Characterization of transcription-deficient temperature-sensitive mutants of herpes simplex virus type 1. Virology. 1978 Dec;91(2):364–379. doi: 10.1016/0042-6822(78)90384-7. [DOI] [PubMed] [Google Scholar]
  44. Watson R. J., Preston C. M., Clements J. B. Separation and characterization of herpes simplex virus type 1 immediate-early mRNA's. J Virol. 1979 Jul;31(1):42–52. doi: 10.1128/jvi.31.1.42-52.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Wigler M., Silverstein S., Lee L. S., Pellicer A., Cheng Y. c., Axel R. Transfer of purified herpes virus thymidine kinase gene to cultured mouse cells. Cell. 1977 May;11(1):223–232. doi: 10.1016/0092-8674(77)90333-6. [DOI] [PubMed] [Google Scholar]
  46. Wigler M., Sweet R., Sim G. K., Wold B., Pellicer A., Lacy E., Maniatis T., Silverstein S., Axel R. Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell. 1979 Apr;16(4):777–785. doi: 10.1016/0092-8674(79)90093-x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES