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. 1984 May;50(2):325–334. doi: 10.1128/jvi.50.2.325-334.1984

Revertants and partial transformants of rat fibroblasts infected with Fujinami sarcoma virus.

B Mathey-Prevot, M Shibuya, J Samarut, H Hanafusa
PMCID: PMC255624  PMID: 6323733

Abstract

Fifteen revertants were isolated from three independent clones of rat fibroblasts transformed by Fujinami sarcoma virus (FSV). Three revertant clones resulted from the deletion of the one copy of the FSV provirus, and one encoded an enzymatically inactive, transformation-defective protein. The remaining revertant clones were characterized by a transcriptional block of the provirus. Digestion of chromosomal DNA with MspI and HpaII revealed that the FSV provirus was hypermethylated in these revertants, whereas proviral DNA of their spontaneous retransformants was hypomethylated. Furthermore, the revertants had lost DNase I-hypersensitive sites in and around the FSV provirus. The effect of transcriptional regulation of the FSV provirus was further analyzed in clones showing various degrees of phenotypic transformation. We quantitated v-fps mRNA levels in these cells by liquid hybridization and found that increasing levels of viral RNA correlated with a more pronounced transformed phenotype. These results suggest that transcription of FSV proviral DNA is under both viral and cellular control and that transformation by FSV is a function of the dosage of v-fps mRNA.

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Selected References

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

  1. Adkins B., Hunter T., Sefton B. M. The transforming proteins of PRCII virus and Rous sarcoma virus form a complex with the same two cellular phosphoproteins. J Virol. 1982 Aug;43(2):448–455. doi: 10.1128/jvi.43.2.448-455.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barbacid M., Donner L., Ruscetti S. K., Sherr C. J. Transformation-defective mutants of Snyder-Theilen feline sarcoma virus lack tyrosine-specific protein kinase activity. J Virol. 1981 Jul;39(1):246–254. doi: 10.1128/jvi.39.1.246-254.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boettiger D. Virogenic nontransformed cells isolated following infection of normal rat kidney cells with B77 strain Rous sarcoma virus. Cell. 1974 Sep;3(1):71–76. doi: 10.1016/0092-8674(74)90042-7. [DOI] [PubMed] [Google Scholar]
  4. Brugge J. S., Erikson E., Erikson R. L. The specific interaction of the Rous sarcoma virus transforming protein, pp60src, with two cellular proteins. Cell. 1981 Aug;25(2):363–372. doi: 10.1016/0092-8674(81)90055-6. [DOI] [PubMed] [Google Scholar]
  5. Brugge J., Yonemoto W., Darrow D. Interaction between the Rous sarcoma virus transforming protein and two cellular phosphoproteins: analysis of the turnover and distribution of this complex. Mol Cell Biol. 1983 Jan;3(1):9–19. doi: 10.1128/mcb.3.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chiswell D. J., Enrietto P. J., Evans S., Quade K., Wyke J. A. Molecular mechanisms involved in morphological variation of avian sarcoma virus-infected rat cells. Virology. 1982 Jan 30;116(2):428–440. doi: 10.1016/0042-6822(82)90137-4. [DOI] [PubMed] [Google Scholar]
  7. Chiswell D. J., Gillespie D. A., Wyke J. A. The changes in proviral chromatin that accompany morphological variation in avian sarcoma virus-infected rat cells. Nucleic Acids Res. 1982 Jul 10;10(13):3967–3980. doi: 10.1093/nar/10.13.3967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Conklin K. F., Coffin J. M., Robinson H. L., Groudine M., Eisenman R. Role of methylation in the induced and spontaneous expression of the avian endogenous virus ev-1: DNA structure and gene products. Mol Cell Biol. 1982 Jun;2(6):638–652. doi: 10.1128/mcb.2.6.638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Courtneidge S. A., Bishop J. M. Transit of pp60v-src to the plasma membrane. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7117–7121. doi: 10.1073/pnas.79.23.7117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Donner L., Turek L. P., Ruscetti S. K., Fedele L. A., Sherr C. J. Transformation-defective mutants of feline sarcoma virus which express a product of the viral src gene. J Virol. 1980 Jul;35(1):129–140. doi: 10.1128/jvi.35.1.129-140.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Drohan W. N., Benade L. E., Graham D. E., Smith G. H. Mouse mammary tumor virus proviral sequences congenital to C3H/Sm mice are differentially hypomethylated in chemically induced, virus-induced, and spontaneous mammary tumors. J Virol. 1982 Sep;43(3):876–884. doi: 10.1128/jvi.43.3.876-884.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Feldman R. A., Hanafusa T., Hanafusa H. Characterization of protein kinase activity associated with the transforming gene product of Fujinami sarcoma virus. Cell. 1980 Dec;22(3):757–765. doi: 10.1016/0092-8674(80)90552-8. [DOI] [PubMed] [Google Scholar]
  13. Gautsch J. W., Wilson M. C. Delayed de novo methylation in teratocarcinoma suggests additional tissue-specific mechanisms for controlling gene expression. Nature. 1983 Jan 6;301(5895):32–37. doi: 10.1038/301032a0. [DOI] [PubMed] [Google Scholar]
  14. Groffen J., Heisterkamp N., Blennerhassett G., Stephenson J. R. Regulation of viral and cellular oncogene expression by cytosine methylation. Virology. 1983 Apr 15;126(1):213–227. doi: 10.1016/0042-6822(83)90473-7. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Haigh L. S., Owens B. B., Hellewell O. S., Ingram V. M. DNA methylation in chicken alpha-globin gene expression. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5332–5336. doi: 10.1073/pnas.79.17.5332. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hanafusa H. Rapid transformation of cells by Rous sarcoma virus. Proc Natl Acad Sci U S A. 1969 Jun;63(2):318–325. doi: 10.1073/pnas.63.2.318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hanafusa T., Mathey-Prevot B., Feldman R. A., Hanafusa H. Mutants of Fujinami sarcoma virus which are temperature sensitive for cellular transformation and protein kinase activity. J Virol. 1981 Apr;38(1):347–355. doi: 10.1128/jvi.38.1.347-355.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hanafusa T., Wang L. H., Anderson S. M., Karess R. E., Hayward W. S., Hanafusa H. Characterization of the transforming gene of Fujinami sarcoma virus. Proc Natl Acad Sci U S A. 1980 May;77(5):3009–3013. doi: 10.1073/pnas.77.5.3009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hayward W. S., Neel B. G., Astrin S. M. Activation of a cellular onc gene by promoter insertion in ALV-induced lymphoid leukosis. Nature. 1981 Apr 9;290(5806):475–480. doi: 10.1038/290475a0. [DOI] [PubMed] [Google Scholar]
  21. Hoffmann J. W., Steffen D., Gusella J., Tabin C., Bird S., Cowing D., Weinberg R. A. DNA methylation affecting the expression of murine leukemia proviruses. J Virol. 1982 Oct;44(1):144–157. doi: 10.1128/jvi.44.1.144-157.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lau A. F., Krzyzek R. A., Faras A. J. Loss of tumorigenicity correlates with a reduction in pp60src kinase activity in a revertant subclone of avian sarcoma virus-infected field vole cells. Cell. 1981 Mar;23(3):815–823. doi: 10.1016/0092-8674(81)90446-3. [DOI] [PubMed] [Google Scholar]
  23. Lee W. H., Bister K., Pawson A., Robins T., Moscovici C., Duesberg P. H. Fujinami sarcoma virus: an avian RNA tumor virus with a unique transforming gene. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2018–2022. doi: 10.1073/pnas.77.4.2018. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lipsich L. A., Cutt J. R., Brugge J. S. Association of the transforming proteins of Rous, Fujinami, and Y73 avian sarcoma viruses with the same two cellular proteins. Mol Cell Biol. 1982 Jul;2(7):875–880. doi: 10.1128/mcb.2.7.875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. Mathey-Prevot B., Hanafusa H., Kawai S. A cellular protein is immunologically crossreactive with and functionally homologous to the Fujinami sarcoma virus transforming protein. Cell. 1982 Apr;28(4):897–906. doi: 10.1016/0092-8674(82)90069-1. [DOI] [PubMed] [Google Scholar]
  28. McGhee J. D., Ginder G. D. Specific DNA methylation sites in the vicinity of the chicken beta-globin genes. Nature. 1979 Aug 2;280(5721):419–420. doi: 10.1038/280419a0. [DOI] [PubMed] [Google Scholar]
  29. McGhee J. D., Wood W. I., Dolan M., Engel J. D., Felsenfeld G. A 200 base pair region at the 5' end of the chicken adult beta-globin gene is accessible to nuclease digestion. Cell. 1981 Nov;27(1 Pt 2):45–55. doi: 10.1016/0092-8674(81)90359-7. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Nomura S., Fischinger P. J., Mattern C. F., Gerwin B. I., Dunn K. J. Revertants of mouse cells transformed by murine sarcoma virus. II. Flat variants induced by fluordeoxyuridine and colcemid. Virology. 1973 Nov;56(1):152–163. doi: 10.1016/0042-6822(73)90294-8. [DOI] [PubMed] [Google Scholar]
  32. Oppermann H., Levinson A. D., Varmus H. E. The structure and protein kinase activity of proteins encoded by nonconditional mutants and back mutants in the sec gene of avian sarcoma virus. Virology. 1981 Jan 15;108(1):47–70. doi: 10.1016/0042-6822(81)90526-2. [DOI] [PubMed] [Google Scholar]
  33. Porzig K. J., Robbins K. C., Aaronson S. A. Cellular regulation of mammalian sarcoma virus expression: a gene regulation model for oncogenesis. Cell. 1979 Apr;16(4):875–884. doi: 10.1016/0092-8674(79)90102-8. [DOI] [PubMed] [Google Scholar]
  34. Quade K., Saule S., Stéhelin D., Kitchener G., Hayman M. J. Revertants of rats cells transformed by avian erythroblastosis virus. Virology. 1981 Dec;115(2):322–333. doi: 10.1016/0042-6822(81)90114-8. [DOI] [PubMed] [Google Scholar]
  35. Razin A., Riggs A. D. DNA methylation and gene function. Science. 1980 Nov 7;210(4470):604–610. doi: 10.1126/science.6254144. [DOI] [PubMed] [Google Scholar]
  36. Shibuya M., Hanafusa H., Balduzzi P. C. Cellular sequences related to three new onc genes of avian sarcoma virus (fps, yes, and ros) and their expression in normal and transformed cells. J Virol. 1982 Apr;42(1):143–152. doi: 10.1128/jvi.42.1.143-152.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Shibuya M., Hanafusa H. Nucleotide sequence of Fujinami sarcoma virus: evolutionary relationship of its transforming gene with transforming genes of other sarcoma viruses. Cell. 1982 Oct;30(3):787–795. doi: 10.1016/0092-8674(82)90283-5. [DOI] [PubMed] [Google Scholar]
  38. Shibuya M., Hanafusa T., Hanafusa H., Stephenson J. R. Homology exists among the transforming sequences of avian and feline sarcoma viruses. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6536–6540. doi: 10.1073/pnas.77.11.6536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Shibuya M., Wang L. H., Hanafusa H. Molecular cloning of the Fujinami sarcoma virus genome and its comparison with sequences of other related transforming viruses. J Virol. 1982 Jun;42(3):1007–1016. doi: 10.1128/jvi.42.3.1007-1016.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. 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]
  41. 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]
  42. Takeya T., Hanafusa H., Junghans R. P., Ju G., Skalka A. M. Comparison between the viral transforming gene (src) of recovered avian sarcoma virus and its cellular homolog. Mol Cell Biol. 1981 Nov;1(11):1024–1037. doi: 10.1128/mcb.1.11.1024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Temin H. M. Origin of retroviruses from cellular moveable genetic elements. Cell. 1980 Oct;21(3):599–600. doi: 10.1016/0092-8674(80)90420-1. [DOI] [PubMed] [Google Scholar]
  44. Temin H. M. Structure, variation and synthesis of retrovirus long terminal repeat. Cell. 1981 Nov;27(1 Pt 2):1–3. doi: 10.1016/0092-8674(81)90353-6. [DOI] [PubMed] [Google Scholar]
  45. Varmus H. E., Quintrell N., Ortiz S. Retroviruses as mutagens: insertion and excision of a nontransforming provirus alter expression of a resident transforming provirus. Cell. 1981 Jul;25(1):23–36. doi: 10.1016/0092-8674(81)90228-2. [DOI] [PubMed] [Google Scholar]
  46. Varmus H. E., Quintrell N., Wyke J. Revertants of an ASV-transformed rat cell line have lost the complete provius or sustained mutations in src. Virology. 1981 Jan 15;108(1):28–46. doi: 10.1016/0042-6822(81)90525-0. [DOI] [PubMed] [Google Scholar]
  47. Waalwijk C., Flavell R. A. MspI, an isoschizomer of hpaII which cleaves both unmethylated and methylated hpaII sites. Nucleic Acids Res. 1978 Sep;5(9):3231–3236. doi: 10.1093/nar/5.9.3231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Weintraub H., Groudine M. Chromosomal subunits in active genes have an altered conformation. Science. 1976 Sep 3;193(4256):848–856. doi: 10.1126/science.948749. [DOI] [PubMed] [Google Scholar]
  49. Weisbrod S. Active chromatin. Nature. 1982 May 27;297(5864):289–295. doi: 10.1038/297289a0. [DOI] [PubMed] [Google Scholar]

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