Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1980 Jul;35(1):129–140. doi: 10.1128/jvi.35.1.129-140.1980

Transformation-defective mutants of feline sarcoma virus which express a product of the viral src gene.

L Donner, L P Turek, S K Ruscetti, L A Fedele, C J Sherr
PMCID: PMC288788  PMID: 6251261

Abstract

Mink cell cultures infected with the Snyder-Theilen strain of feline sarcoma-leukemia virus were cloned from single cells under conditions favoring single virus-single cell interactions. The primary colonies included (i) typical feline sarcoma virus (FeSV)-transformed nonproducer clones, one of which segregated revertants, and (ii) FeSV-infected, phenotypically normal clones, three of which spontaneously converted to the transformed phenotype. The revertants and spontaneous transformants were compared with parental and sister clones expressing the opposite phenotype. Transformed subclones formed colonies in agar, were tumorigenic in nude mice, and failed to bind epidermal growth factor, whereas flat sister subclones were indistinguishable from uninfected mink cells in each of these assays. Sister subclones derived from the same infectious event contained FeSV proviruses integrated at the same molecular site, regardless of which phenotype was expressed. One revertant clone, however, lacked most FeSV proviral DNA sequences but retained terminal portions of the FeSV genome which persisted at the original site of proviral DNA insertion. Two flat subclones expressed viral RNA and the phosphorylated "gag-x" polyprotein (pp78gag-x) encoded by the gag and src sequences of the FeSV genome. Both of these clones were susceptible to retransformation by FeSV. Although unable to induce foci, the viruses rescued from these cells contained as much FeSV RNA as the focus-forming viruses rescued from transformed sister subclones and could be retransmitted to mink cells, again inducing FeSV gene products without signs of morphological transformation. We conclude that these FeSV genomes represent transformation-defective mutants.

Full text

PDF
129

Images in this article

134Image
on p.134
136Image
on p.136

Selected References

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

  1. Ando T. A nuclease specific for heat-denatured DNA in isolated from a product of Aspergillus oryzae. Biochim Biophys Acta. 1966 Jan 18;114(1):158–168. doi: 10.1016/0005-2787(66)90263-2. [DOI] [PubMed] [Google Scholar]
  2. Barbacid M., Lauver A. V., Devare S. G. Biochemical and immunological characterization of polyproteins coded for by the McDonough, Gardner-Arnstein, and Snyder-Theilen strains of feline sarcoma virus. J Virol. 1980 Jan;33(1):196–207. doi: 10.1128/jvi.33.1.196-207.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bensinger W. I., Robbins K. C., Greenberger J. S., Aaronson S. A. Different mechanisms for morphologic reversion of a clonal population of murine sarcoma virus-transformed nonproducer cells. Virology. 1977 Apr;77(2):750–761. doi: 10.1016/0042-6822(77)90496-2. [DOI] [PubMed] [Google Scholar]
  4. Bister K., Hayman M. J., Vogt P. K. Defectiveness of avian myelocytomatosis virus MC29: isolation of long-term nonproducer cultures and analysis of virus-specific polypeptide synthesis. Virology. 1977 Oct 15;82(2):431–448. doi: 10.1016/0042-6822(77)90017-4. [DOI] [PubMed] [Google Scholar]
  5. Bister K., Löliger H. C., Duesberg P. H. Oligoribonucleotide map and protein of CMII: detection of conserved and nonconserved genetic elements in avian acute leukemia viruses CMII, MC29, and MH2. J Virol. 1979 Oct;32(1):208–219. doi: 10.1128/jvi.32.1.208-219.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Britten R. J., Kohne D. E. Repeated sequences in DNA. Hundreds of thousands of copies of DNA sequences have been incorporated into the genomes of higher organisms. Science. 1968 Aug 9;161(3841):529–540. doi: 10.1126/science.161.3841.529. [DOI] [PubMed] [Google Scholar]
  7. Collett M. S., Brugge J. S., Erikson R. L., Lau A. F., Krzyzek R. A., Faras A. J. The src gene product of transformed and morphologically reverted ASV-infected mammalian cells. Nature. 1979 Sep 20;281(5728):195–198. doi: 10.1038/281195a0. [DOI] [PubMed] [Google Scholar]
  8. Fischinger P. J., Blevins C. S., Nomura S. Simple, quantitative assay for both xenotropic murine leukemia and ecotropic feline leukemia viruses. J Virol. 1974 Jul;14(1):177–179. doi: 10.1128/jvi.14.1.177-179.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fischinger P. J., Nomura S., Tuttle-Fuller N., Dunn K. J. Revertants of mouse cells transformed by murine sarcoma virus. 3. Metastable expression of virus functions in revertants retransformed by murine sarcoma virus. Virology. 1974 May;59(1):217–229. [PubMed] [Google Scholar]
  10. Frankel A. E., Gilbert J. H., Porzig K. J., Scolnick E. M., Aaronson S. A. Nature and distribution of feline sarcoma virus nucleotide sequences. J Virol. 1979 Jun;30(3):821–827. doi: 10.1128/jvi.30.3.821-827.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goldberg R. J., Levin R., Parks W. P., Scolnick E. M. Quantitative analysis of the rescue of RNA sequences by mammalian type C viruses. J Virol. 1975 Jan;17(1):43–50. doi: 10.1128/jvi.17.1.43-50.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Greenberger J. S., Anderson G. R., Aaronson S. A. Transformation-defective virus mutants in a class of morphologic revertants of sarcoma virus transformed nonproducer cells. Cell. 1974 Aug;2(4):279–286. doi: 10.1016/0092-8674(74)90022-1. [DOI] [PubMed] [Google Scholar]
  13. Hayman M. J., Royer-Pokora B., Graf T. Defectiveness of avian erythroblastosis virus: synthesis of a 75K gag-related protein. Virology. 1979 Jan 15;92(1):31–45. doi: 10.1016/0042-6822(79)90212-5. [DOI] [PubMed] [Google Scholar]
  14. Henderson I. C., Lieber M. M., Todaro G. J. Mink cell line Mv 1 Lu (CCL 64). Focus formation and the generation of "nonproducer" transformed cell lines with murine and feline sarcoma viruses. Virology. 1974 Jul;60(1):282–287. doi: 10.1016/0042-6822(74)90386-9. [DOI] [PubMed] [Google Scholar]
  15. Khan A. S., Stephenson J. R. Feline leukemia virus: biochemical and immunological characterization of gag gene-coded structural proteins. J Virol. 1977 Sep;23(3):599–607. doi: 10.1128/jvi.23.3.599-607.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Mellon P., Pawson A., Bister K., Martin G. S., Duesberg P. H. Specific RNA sequences and gene products of MC29 avian acute leukemia virus. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5874–5878. doi: 10.1073/pnas.75.12.5874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Palmiter R. D. Magnesium precipitation of ribonucleoprotein complexes. Expedient techniques for the isolation of undergraded polysomes and messenger ribonucleic acid. Biochemistry. 1974 Aug 13;13(17):3606–3615. doi: 10.1021/bi00714a032. [DOI] [PubMed] [Google Scholar]
  19. Porzig K. J., Barbacid M., Aaronson S. A. Biological properties and translational products of three independent isolates of feline sarcoma virus. Virology. 1979 Jan 15;92(1):91–107. doi: 10.1016/0042-6822(79)90217-4. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. Ringold G., Lasfargues E. Y., Bishop J. M., Varmus H. E. Production of mouse mammary tumor virus by cultured cells in the absence and presence of hormones: assay by molecular hybridization. Virology. 1975 May;65(1):135–147. doi: 10.1016/0042-6822(75)90014-8. [DOI] [PubMed] [Google Scholar]
  23. Robbins K. C., Barbacid M., Porzig K. J., Aaronson S. A. Involvement of different exogenous feline leukemia virus subgroups in the generation of independent feline sarcoma virus isolates. Virology. 1979 Aug;97(1):1–11. doi: 10.1016/0042-6822(79)90367-2. [DOI] [PubMed] [Google Scholar]
  24. Ruscetti S. K., Linemeyer D., Feild J., Troxler D., Scolnick E. M. Characterization of a protein found in cells infected with the spleen focus-forming virus that shares immunological cross-reactivity with the gp70 found in mink cell focus-inducing virus particles. J Virol. 1979 Jun;30(3):787–798. doi: 10.1128/jvi.30.3.787-798.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ruscetti S. K., Turek L. P., Sherr C. J. Three independent isolates of feline sarcoma virus code for three distinct gag-x polyproteins. J Virol. 1980 Jul;35(1):259–264. doi: 10.1128/jvi.35.1.259-264.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sherr C. J., Benveniste R. E., Todaro G. J. Endogenous mink (Mustela vison) type C virus isolated from sarcoma virus-transformed mink cells. J Virol. 1978 Mar;25(3):738–749. doi: 10.1128/jvi.25.3.738-749.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sherr C. J., Fedele L. A., Donner L., Turek L. P. Restriction endonuclease mapping of unintegrated proviral DNA of Snyder-Theilen feline sarcoma virus: localization of sarcoma-specific sequences. J Virol. 1979 Dec;32(3):860–875. doi: 10.1128/jvi.32.3.860-875.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sherr C. J., Fedele L. A., Oskarsson M., Maizel J., Vande Woude G. Molecular cloning of Snyder-Theilen feline leukemia and sarcoma viruses: comparative studies of feline sarcoma virus with its natural helper virus and with Moloney murine sarcoma virus. J Virol. 1980 Apr;34(1):200–212. doi: 10.1128/jvi.34.1.200-212.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sherr C. J., Sen A., Todaro G. J., Sliski A., Essex M. Pseudotypes of feline sarcoma virus contain an 85,000-dalton protein with feline oncornavirus-associated cell membrane antigen (FOCMA) activity. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1505–1509. doi: 10.1073/pnas.75.3.1505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Snyder S. P., Theilen G. H. Transmissible feline fibrosarcoma. Nature. 1969 Mar 15;221(5185):1074–1075. doi: 10.1038/2211074a0. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Stephenson J. R., Khan A. S., Sliski A. H., Essex M. Feline oncornavirus-associated cell membrane antigen: evidence for an immunologically crossreactive feline sarcoma virus-coded protein. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5608–5612. doi: 10.1073/pnas.74.12.5608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Todaro G. J., De Larco J. E., Cohen S. Transformation by murine and feline sarcoma viruses specifically blocks binding of epidermal growth factor to cells. Nature. 1976 Nov 4;264(5581):26–31. doi: 10.1038/264026a0. [DOI] [PubMed] [Google Scholar]
  34. Transplantable murine tumors release mouse-tropic and xenotropic type-C viruses. Int J Cancer. 1975 Apr 15;15(4):555–560. doi: 10.1002/ijc.2910150404. [DOI] [PubMed] [Google Scholar]
  35. Witte O. N., Rosenberg N., Paskind M., Shields A., Baltimore D. Identification of an Abelson murine leukemia virus-encoded protein present in transformed fibroblast and lymphoid cells. Proc Natl Acad Sci U S A. 1978 May;75(5):2488–2492. doi: 10.1073/pnas.75.5.2488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. de Larco J. E., Todaro G. J. Growth factors from murine sarcoma virus-transformed cells. Proc Natl Acad Sci U S A. 1978 Aug;75(8):4001–4005. doi: 10.1073/pnas.75.8.4001. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES