Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1984 Apr;4(4):611–617. doi: 10.1128/mcb.4.4.611

Changes in structure and methylation pattern in a cluster of thymidine kinase genes.

B A Christy, G A Scangos
PMCID: PMC368766  PMID: 6717437

Abstract

Cell line 101 is a thymidine kinase (TK)-positive derivative of Ltk- which contains ca. 20 copies of the herpes simplex virus TK gene organized in a tandem array. DNA methylation at three sites within the gene and flanking sequences was inversely correlated with expression: the sites were unmethylated in line 101, methylated in each of 4 TK-negative derivatives of 101, and unmethylated in each of 21 TK-positive derivatives derived from them. The same three sites were affected in most of the 20 copies of the TK gene, whereas other sites between them were not affected. Although the entire gene cluster was never lost, indicating that integration into the genome was stable, internal rearrangements occurred at a high frequency. The rearrangements had no obvious correlation with the state of methylation or with the expression of the genes.

Full text

PDF
611

Images in this article

613Image
on p.613
613Image
on p.613
614Image
on p.614
614Image
on p.614
615Image
on p.615

Selected References

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

  1. Busslinger M., Hurst J., Flavell R. A. DNA methylation and the regulation of globin gene expression. Cell. 1983 Aug;34(1):197–206. doi: 10.1016/0092-8674(83)90150-2. [DOI] [PubMed] [Google Scholar]
  2. Charache S., Dover G., Smith K., Talbot C. C., Jr, Moyer M., Boyer S. Treatment of sickle cell anemia with 5-azacytidine results in increased fetal hemoglobin production and is associated with nonrandom hypomethylation of DNA around the gamma-delta-beta-globin gene complex. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4842–4846. doi: 10.1073/pnas.80.15.4842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Christy B., Scangos G. Expression of transferred thymidine kinase genes is controlled by methylation. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6299–6303. doi: 10.1073/pnas.79.20.6299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clough D. W., Kunkel L. M., Davidson R. L. 5-Azacytidine-induced reactivation of a herpes simplex thymidine kinase gene. Science. 1982 Apr 2;216(4541):70–73. doi: 10.1126/science.6175023. [DOI] [PubMed] [Google Scholar]
  5. Davies R. L., Fuhrer-Krusi S., Kucherlapati R. S. Modulation of transfected gene expression mediated by changes in chromatin structure. Cell. 1982 Dec;31(3 Pt 2):521–529. doi: 10.1016/0092-8674(82)90308-7. [DOI] [PubMed] [Google Scholar]
  6. Elgin S. C. DNAase I-hypersensitive sites of chromatin. Cell. 1981 Dec;27(3 Pt 2):413–415. doi: 10.1016/0092-8674(81)90381-0. [DOI] [PubMed] [Google Scholar]
  7. Enquist L. W., Vande Woude G. F., Wagner M., Smiley J. R., Summers W. C. Construction and characterization of a recombinant plasmid encoding the gene for the thymidine kinase of Herpes simplex type 1 virus. Gene. 1979 Nov;7(3-4):335–342. doi: 10.1016/0378-1119(79)90052-0. [DOI] [PubMed] [Google Scholar]
  8. Gerber-Huber S., May F. E., Westley B. R., Felber B. K., Hosbach H. A., Andres A. C., Ryffel G. U. In contrast to other Xenopus genes the estrogen-inducible vitellogenin genes are expressed when totally methylated. Cell. 1983 May;33(1):43–51. doi: 10.1016/0092-8674(83)90333-1. [DOI] [PubMed] [Google Scholar]
  9. Groudine M., Eisenman R., Weintraub H. Chromatin structure of endogenous retroviral genes and activation by an inhibitor of DNA methylation. Nature. 1981 Jul 23;292(5821):311–317. doi: 10.1038/292311a0. [DOI] [PubMed] [Google Scholar]
  10. Guerry P., LeBlanc D. J., Falkow S. General method for the isolation of plasmid deoxyribonucleic acid. J Bacteriol. 1973 Nov;116(2):1064–1066. doi: 10.1128/jb.116.2.1064-1066.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Harbers K., Schnieke A., Stuhlmann H., Jähner D., Jaenisch R. DNA methylation and gene expression: endogenous retroviral genome becomes infectious after molecular cloning. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7609–7613. doi: 10.1073/pnas.78.12.7609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hardies S. C., Axelrod D. E., Edgell M. H., Hutchison C. A., 3rd Phenotypic variation associated with molecular alterations at a cluster of thymidine kinase genes. Mol Cell Biol. 1983 Jul;3(7):1163–1171. doi: 10.1128/mcb.3.7.1163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Huttner K. M., Scangos G. A., Ruddle F. H. DNA-mediated gene transfer of a circular plasmid into murine cells. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5820–5824. doi: 10.1073/pnas.76.11.5820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Lewis J. A., Mengheri E., Esteban M. Induction of an antiviral response by interferon requires thymidine kinase. Proc Natl Acad Sci U S A. 1983 Jan;80(1):26–30. doi: 10.1073/pnas.80.1.26. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mandel J. L., Chambon P. DNA methylation: organ specific variations in the methylation pattern within and around ovalbumin and other chicken genes. Nucleic Acids Res. 1979 Dec 20;7(8):2081–2103. doi: 10.1093/nar/7.8.2081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McGeady M. L., Jhappan C., Ascione R., Vande Woude G. F. In vitro methylation of specific regions of the cloned Moloney sarcoma virus genome inhibits its transforming activity. Mol Cell Biol. 1983 Mar;3(3):305–314. doi: 10.1128/mcb.3.3.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. McKeon C., Ohkubo H., Pastan I., de Crombrugghe B. Unusual methylation pattern of the alpha 2 (l) collagen gene. Cell. 1982 May;29(1):203–210. doi: 10.1016/0092-8674(82)90104-0. [DOI] [PubMed] [Google Scholar]
  20. Niwa O., Sugahara T. 5-Azacytidine induction of mouse endogenous type C virus and suppression of DNA methylation. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6290–6294. doi: 10.1073/pnas.78.10.6290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ostrander M., Vogel S., Silverstein S. Phenotypic switching in cells transformed with the herpes simplex virus thymidine kinase gene. Mol Cell Biol. 1982 Jun;2(6):708–714. doi: 10.1128/mcb.2.6.708. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ott M. O., Sperling L., Cassio D., Levilliers J., Sala-Trepat J., Weiss M. C. Undermethylation at the 5' end of the albumin gene is necessary but not sufficient for albumin production by rat hepatoma cells in culture. Cell. 1982 Oct;30(3):825–833. doi: 10.1016/0092-8674(82)90287-2. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Robins D. M., Axel R., Henderson A. S. Chromosome structure and DNA sequence alterations associated with mutation of transformed genes. J Mol Appl Genet. 1981;1(3):191–203. [PubMed] [Google Scholar]
  25. Simon D., Stuhlmann H., Jähner D., Wagner H., Werner E., Jaenisch R. Retrovirus genomes methylated by mammalian but not bacterial methylase are non-infectious. Nature. 1983 Jul 21;304(5923):275–277. doi: 10.1038/304275a0. [DOI] [PubMed] [Google Scholar]
  26. Stein R., Sciaky-Gallili N., Razin A., Cedar H. Pattern of methylation of two genes coding for housekeeping functions. Proc Natl Acad Sci U S A. 1983 May;80(9):2422–2426. doi: 10.1073/pnas.80.9.2422. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stuhlmann H., Jähner D., Jaenisch R. Infectivity and methylation of retroviral genomes is correlated with expression in the animal. Cell. 1981 Oct;26(2 Pt 2):221–232. doi: 10.1016/0092-8674(81)90305-6. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Sweet R. W., Chao M. V., Axel R. The structure of the thymidine kinase gene promoter: nuclease hypersensitivity correlates with expression. Cell. 1982 Dec;31(2 Pt 1):347–353. doi: 10.1016/0092-8674(82)90128-3. [DOI] [PubMed] [Google Scholar]
  30. Vardimon L., Neumann R., Kuhlmann I., Sutter D., Doerfler W. DNA methylation and viral gene expression in adenovirus-transformed and -infected cells. Nucleic Acids Res. 1980 Jun 11;8(11):2461–2473. doi: 10.1093/nar/8.11.2461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Waechter D. E., Baserga R. Effect of methylation on expression of microinjected genes. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1106–1110. doi: 10.1073/pnas.79.4.1106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wigler M., Levy D., Perucho M. The somatic replication of DNA methylation. Cell. 1981 Apr;24(1):33–40. doi: 10.1016/0092-8674(81)90498-0. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. van der Ploeg L. H., Flavell R. A. DNA methylation in the human gamma delta beta-globin locus in erythroid and nonerythroid tissues. Cell. 1980 Apr;19(4):947–958. doi: 10.1016/0092-8674(80)90086-0. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES