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Journal of General Virology header logo Journal of General Virology

Volume 62, Issue 2

Transfer of Murine Leukaemia and Murine Sarcoma Virus Genetic Information by Transfection with Isolated Metaphase Chromosomes No Access

Abstract

SUMMARY

Chromosome-mediated transfer of murine leukaemia (MuLV) and murine sarcoma (MuSV) virus genetic information to uninfected recipient cells was investigated. Metaphase chromosomes from AKR MuLV-infected SC-1 mouse cells were incubated with NIH/3T3 cells. After several passages (1 to 3 weeks), infectious virions exhibiting reverse transcriptase activity and the characteristic host range of ecotropic, N-tropic AKR virus appeared in the supernatant fluids of the treated cells. Restriction endonuclease analysis of genomic DNA from transfected cells indicated that AKR proviral DNA was associated with the high molecular weight DNA of the host. These results demonstrate that the AKR MuLV genome can be stably transferred to uninfected recipient cells via isolated metaphase chromosomes. Although AKR virions are not able to infect heterologous cells, chromosome-mediated transfection resulted in the establishment of productive AKR MuLV infection in mink cells. Thus, the use of chromosomes to transfer virus genes can circumvent the natural host restriction barrier. In other experiments, it was shown that normal NIH/3T3 cells were transformed after exposure to metaphase chromosomes isolated from an MuSV-infected, non-producer line. Foci were detected 14 to 21 days after chromosome treatment and were shown to contain true viral transformants since transforming virus was produced after superinfection with MuLV.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): gene transfer , metaphase chromosomes , murine retroviruses and transformation
© Society for General Microbiology 1982
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1982-10-01
2025-05-16
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References

  1. Aaronson S. A., Dunn C. Y. 1974; High-frequency C-type virus induction by inhibitors of protein synthesis. Science 183:422–424
     [Google Scholar]
  2. Benjers B. M., Bassin R. H., Rein A., Gerwin B. I., Duran-Troise G. 1979; Mechanism of restriction of murine leukemia viruses varies between different strains of Fv-ln mice. International Journal of Cancer 24:600–607
     [Google Scholar]
  3. Besmer P., Baltimore D. 1977; Mechanism of restriction of ecotropic and xenotropic murine leukemia viruses and formation of pseudotypes between the two viruses. Journal of Virology 21:965–973
     [Google Scholar]
  4. Burch J. W., McBride O. W. 1975; Human gene expression in rodent cells after uptake of isolated metaphase chromosomes. Proceedings of the National Academy of Sciences of the United States of America 72:1797–1801
     [Google Scholar]
  5. Burkholder G., Mukherjee B. B. 1970; Uptake of isolated metaphase chromosomes by mammalian cells in vitro. Experimental Cell Research 61:413–418
     [Google Scholar]
  6. Carrano A. V., Gray J. W., Langlois R. G., Burkhart Schultz K. J., Van Dilla M. A. 1979; Measurement and purification of human chromosomes by flow cytometry and sorting. Proceedings of the National Academy of Sciences of the United States of America 76:1382–1384
     [Google Scholar]
  7. Cassingena R., Suarez H. G., Lavialle C., Persuy M. A., Ermonval M. 1978; Transformation of normal diploid cells by isolated metaphase chromosomes of virus-transformed or spontaneous tumor cells. Gene 4:337–349
     [Google Scholar]
  8. Chattopadhyay S. K., Cloyd M. W., Linemeyer D. L., Lander M. R., Rands E., Lowy D. R. 1982; Cellular origin and role of mink cell focus-forming viruses in murine thymic lymphomas. Nature, London 295:25–31
     [Google Scholar]
  9. Dhar R., Mcclements W. L., Enquist L. W., Vande Woude G. F. 1980; Nucleotide sequences of integrated Moloney sarcoma provirus long terminal repeats and their host and viral junctions. Proceedings oj the National Academy of Sciences of the United States of America 77:3937–3941
     [Google Scholar]
  10. Ebina T., Miao R., Watanabe Y. 1974; Transfer of chromosomal material: association of rescuable sarcoma virus genome with the chromosomal fraction. Experimental Cell Research 88:203–206
     [Google Scholar]
  11. Gisselbrecht S., Bassin R. H., Gerwin B. I., Rein A. 1974; Dual susceptibility of a 3T3 mouse cell line to infection by N- and B-tropic murine leukemia virus: apparent lack of expression of the Fv-1 gene. International Journal of Cancer 14:106–113
     [Google Scholar]
  12. Hager G. L., Chang E. H., Chan H. W., Garon C. F., Israel M. A., Martin M. A., Scolnick E. M., Lowy D. R. 1979; Molecular cloning of the Harvey sarcoma virus closed circular DNA intermediates: initial structural and biological characterization. Journal of Virology 31:795–809
     [Google Scholar]
  13. Hartley J. W., Rowe W. P. 1975; Clonal cell lines from a feral mouse embryo which lack host-range restrictions for murine leukemia viruses. Virology 65:128–134
     [Google Scholar]
  14. Hartley J. W., Rowe W. P., Huebner R. J. 1970; Host-range restrictions of murine leukemia viruses in mouse embryo cell cultures. Journal of Virology 5:221–225
     [Google Scholar]
  15. Hill M., Hillova J. 1972; Virus recovery in chicken cells tested with Rous sarcoma cell DNA. Nature New Biology 237:35–39
     [Google Scholar]
  16. Igo-Kemenes T., Horz W., Zachau H. G. 1982; Chromatin. Annual Review of Biochemistry 51:89–121
     [Google Scholar]
  17. Ishimoto A., Hartley J. W., Rowe W. P. 1978; Phenotypic mixing between murine leukemia viruses: characteristics of ecotropic virus infection of heterologous cells. Virology 91:464–471
     [Google Scholar]
  18. Klobutcher L. A., Ruddle F. H. 1979; Phenotype stabilization and integration of transferred material in chromosome-mediated gene transfer. Nature, London 280:657–660
     [Google Scholar]
  19. Levin J. G., Rosenak M. J. 1976; Synthesis of murine leukemia virus proteins associated with virions assembled in actinomycin D-treated cells: evidence for persistence of viral messenger RNA. Proceedings oj the National Academy of Sciences of the United States of America 73:1154–1158
     [Google Scholar]
  20. Lewis W. H., Srinivasan P. R., Stokoe N., Siminovitch L. 1980; Parameters governing the transfer of the genes for thymidine kinase and dihydrofolate reductase into mouse cells using metaphase chromosomes or DNA. Somatic Cell Genetics 6:333–347
     [Google Scholar]
  21. Lowy D. R., Rands E., Scolnick E. M. 1978; Helper-independent transformation by unintegrated Harvey sarcoma virus DNA. Journal of Virology 26:291–298
     [Google Scholar]
  22. Lowy D. R., Rands E., Chattopadhyay S. K., Garon C. F., Hager G. L. 1980; Molecular cloning of infectious integrated murine leukemia virus DNA from infected mouse cells. Proceedings of the National Academy of Sciences of the United States of America 77:614–618
     [Google Scholar]
  23. Lueders K., Leder A., Leder P., Kuff E. 1982; Association between a transposed a giobin pseudogene and retrovirus-like elements in the Balb/c mouse genome. Nature, London 295:426–428
     [Google Scholar]
  24. McBride O. W., Ozer H. L. 1973; Transfer of genetic information by purified metaphase chromosomes. Proceedings of the National Academy of Sciences of the United States of America 70:1258–1262
     [Google Scholar]
  25. Mamaus T., Jeffrey A., Kleid D. G. 1975; Nucleotide sequence of the rightward operator of phage 2. Proceedings of the National Academy of Sciences of the United States of America 72:1184–1188
     [Google Scholar]
  26. Miller C. L., Ruddle F. H. 1978; Co-transfer of human X-linked markers into murine somatic cells via isolated metaphase chromosomes. Proceedings of the National Academy of Sciences of the United States of America 74:3346–3350
     [Google Scholar]
  27. Mukherjee A. B., Orloff S., Butler J. Deb., Triche T., Lalley P., Schulman J. D. 1978; Entrapment of metaphase chromosomes into phospholipid vesicles (lipochromosomes): carrier potential in gene transfer. Proceedings of the National Academy of Sciences of the United States of America 75:1361–1365
     [Google Scholar]
  28. Neel B. G., Hayward W. S., Robinson H. L., Fang J., Astrin S. M. 1981; Avian leukosis virus-induced tumors have common proviral integration sites and synthesize discrete new RNAs: oncogenesis by promoter insertion. Cell 23:323–334
     [Google Scholar]
  29. Padgett T. G., Stubblefield E., Varmus H. E. 1977; Chicken macrochromosomes contain an endogenous provirus and microchromosomes contain sequences related to the transforming gene of ASV. Cell 10:649–657
     [Google Scholar]
  30. Payne G. S., Courtneidge S. A., Crittenden L. B., Fadly A. M., Bishop J. M., Varmus H. E. 1981; Analysis of avian leukosis virus DNA and RNA in bursal tumors: viral gene expression is not required for maintenance of the tumor state. Cell 23:311–322
     [Google Scholar]
  31. Pincus T., Rowe W. P., Lilly F. 1971; A major genetic locus affecting resistance to infection with murine leukemia viruses. II. Apparent identity to a major locus described for resistance to Friend murine leukemia virus. Journal of Experimental Medicine 133:1234–1241
     [Google Scholar]
  32. Rands E., Lowy D. R., Lander M. R., Chattopadhyay S. K. 1981; Restriction endonuclease mapping of ecotropic murine leukemia virus DNAs: size and sequence heterogeneity of the long terminal repeat. Virology 108:445–452
     [Google Scholar]
  33. Rowe W. P., Pugh W. E., Hartley J. W. 1970; Plaque assay techniques for murine leukemia viruses. Virology 42:1136–1139
     [Google Scholar]
  34. Shani M., Huberman E., Aloni Y., Sachs L. 1974; Activation of simian virus 40 by transfer of isolated chromosomes from transformed cells. Virology 61:303–305
     [Google Scholar]
  35. Shih C., Shilo B. Z., Goldfarb M. P., Dannenberg A., Weinberg R. A. 1979; Passage of phenotypes of chemically transformed cells via transfection of DNA and chromatin. Proceedings of the National Academy of Sciences of the United States of America 76:5714–5718
     [Google Scholar]
  36. Shimotohno K., Mizutanl S., Temin H. M. 1980; Sequence of retrovirus provirus resembles that of bacterial transposable elements. Nature, London 285:550–554
     [Google Scholar]
  37. Shoemaker C., Goff S., Gilboa E., Paskind M., Mitra S. W., Baltimore D. 1980; Structure of a cloned circular Moloney murine leukemia virus DNA molecule containing an inverted segment: implications for retrovirus integration. Proceedings of the National Academy of Sciences of the United States of America 77:3932–3936
     [Google Scholar]
  38. Southern E. M. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98:503–517
     [Google Scholar]
  39. Steffen D., Bird S., Rowe W. P., Weinberg R. A. 1979; Identification of DNA fragments carrying ecotropic proviruses of AKR mice. Proceedings of the National Academy of Sciences of the United States of America 76:4554–4558
     [Google Scholar]
  40. Sutcliffe J. G., Shinnick T. M., Verma I. M., Lerner R. A. 1980; Nucleotide sequence of Moloney leukemia virus: 3′ end reveals details of replication, analogy to bacterial transposons, and an unexpected gene. Proceedings of the National Academy of Sciences of the United States of America 77:3302–3306
     [Google Scholar]
  41. Tereba A., Lai M. M. C., Murti K. G. 1979; Chromosome 1 contains the endogenous RAV-0 retrovirus sequences in chicken cells. Proceedings of the National Academy of Sciences of the United States of America 76:6486–6490
     [Google Scholar]
  42. Van Beveren C., Goddard J. G., Berns A., Verma I. M. 1980; Structure of Moloney murine leukemia viral DNA: nucleotide sequence of the 5′ long terminal repeat and adjacent cellular sequences. Proceedings of the National Academy of Sciences of the United States of America 77:3307–3311
     [Google Scholar]
  43. Willecke K., Ruddle F. H. 1975; Transfer of the human gene for hypoxanthine-guanine phosphoribosyl-transferase via isolated human metaphase chromosomes into mouse L-cells. Proceedings of the National Academy of Sciences of the United States of America 72:1792–1796
     [Google Scholar]
/content/journal/jgv/10.1099/0022-1317-62-2-227
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Transfer of Murine Leukaemia and Murine Sarcoma Virus Genetic Information by Transfection with Isolated Metaphase Chromosomes
J. Gen. Virol. 62, 227 (1982); https://doi.org/10.1099/0022-1317-62-2-227
/content/journal/jgv/10.1099/0022-1317-62-2-227
/content/journal/jgv/10.1099/0022-1317-62-2-227
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