Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1987 Mar;61(3):889–897. doi: 10.1128/jvi.61.3.889-897.1987

A temperature-sensitive mutant of the myeloproliferative sarcoma virus, altered by a point mutation in the mos oncogene, has been modified as a selectable retroviral vector.

J Friel, C Stocking, A Stacey, W Ostertag
PMCID: PMC254034  PMID: 3027415

Abstract

The myeloproliferative sarcoma virus (MPSV) is a mos-oncogenic retrovirus which induces an acute myeloproliferative disease in adult mice. The isolation and molecular cloning of two mutants of MPSV temperature sensitive (ts) for mos transformation (Kollek et al., J. Virol. 50:717-724, 1984) have been described previously. In this report, we describe the biological activity of these clones, the molecular basis of the ts lesion of one clone, and the construction of a selectable vector based on the MPSV ts genome. Both molecular clones, ts159 and ts124, proved to have retained the ts phenotype, the former being tighter for the induction and maintenance of the transformed phenotype. A single transition (G----A) at position 1888 in the mos coding region, resulting in the change of Gly to Arg at position 307, was responsible for the ts phenotype of clone ts159. Substitution of sequences carrying this mutation with the corresponding sequences of the wild-type virus generated a virus that was ts for transformation. Insertion of the dominant selectable marker gene for geneticin resistance (neor) into ts159 did not disrupt mos expression or its ts phenotype. neor-ts159 facilitates the study of mos action by allowing the selection of infected cells at the nonpermissive temperature before mos transformation has been induced. Furthermore, infected cells which show no obvious phenotype alteration due to mos expression can be identified by their Neor phenotype.

Full text

PDF
889

Selected References

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

  1. Arlinghaus R. B. The ts110 Moloney mouse sarcoma virus system: gag-mos gene products and cellular transformation. J Gen Virol. 1985 Sep;66(Pt 9):1845–1853. doi: 10.1099/0022-1317-66-9-1845. [DOI] [PubMed] [Google Scholar]
  2. Barker W. C., Dayhoff M. O. Viral src gene products are related to the catalytic chain of mammalian cAMP-dependent protein kinase. Proc Natl Acad Sci U S A. 1982 May;79(9):2836–2839. doi: 10.1073/pnas.79.9.2836. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bishop J. M. Viral oncogenes. Cell. 1985 Aug;42(1):23–38. doi: 10.1016/s0092-8674(85)80098-2. [DOI] [PubMed] [Google Scholar]
  4. Bister K., Jansen H. W. Oncogenes in retroviruses and cells: biochemistry and molecular genetics. Adv Cancer Res. 1986;47:99–188. doi: 10.1016/s0065-230x(08)60199-2. [DOI] [PubMed] [Google Scholar]
  5. Blair D. G., Hull M. A., Finch E. A. The isolation and preliminary characterization of temperature-sensitive transformation mutants of Moloney sarcoma virus. Virology. 1979 Jun;95(2):303–316. doi: 10.1016/0042-6822(79)90486-0. [DOI] [PubMed] [Google Scholar]
  6. Blair D. G., McClements W. L., Oskarsson M. K., Fischinger P. J., Vande Woude G. F. Biological activity of cloned Moloney sarcoma virus DNA: Terminally redundant sequences may enhance transformation efficiency. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3504–3508. doi: 10.1073/pnas.77.6.3504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bold R. J., Donoghue D. J. Biologically active mutants with deletions in the v-mos oncogene assayed with retroviral vectors. Mol Cell Biol. 1985 Nov;5(11):3131–3138. doi: 10.1128/mcb.5.11.3131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Canaani E., Robbins K. C., Aaronson S. A. The transforming gene of Moloney murine sarcoma virus. Nature. 1979 Nov 22;282(5737):378–383. doi: 10.1038/282378a0. [DOI] [PubMed] [Google Scholar]
  10. Cizdziel P. E., Nash M. A., Blair D. G., Murphy E. C., Jr Molecular basis underlying phenotypic revertants of Moloney murine sarcoma virus MuSVts110. J Virol. 1986 Jan;57(1):310–317. doi: 10.1128/jvi.57.1.310-317.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Colbère-Garapin F., Horodniceanu F., Kourilsky P., Garapin A. C. A new dominant hybrid selective marker for higher eukaryotic cells. J Mol Biol. 1981 Jul 25;150(1):1–14. doi: 10.1016/0022-2836(81)90321-1. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Duesberg P. H. Activated proto-onc genes: sufficient or necessary for cancer? Science. 1985 May 10;228(4700):669–677. doi: 10.1126/science.3992240. [DOI] [PubMed] [Google Scholar]
  14. Fagg B., Ostertag W., Klein B., Le Bousse C. Myeloproliferative sarcoma virus: its effects on erythropoiesis in adult DBA/2J mice. J Cell Physiol. 1983 Jul;116(1):16–20. doi: 10.1002/jcp.1041160104. [DOI] [PubMed] [Google Scholar]
  15. Frankel A. E., Fischinger P. J. Nucleotide sequences in mouse DNA and RNA specific for Moloney sarcoma virus. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3705–3709. doi: 10.1073/pnas.73.10.3705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Franz T., Hilberg F., Seliger B., Stocking C., Ostertag W. Retroviral mutants efficiently expressed in embryonal carcinoma cells. Proc Natl Acad Sci U S A. 1986 May;83(10):3292–3296. doi: 10.1073/pnas.83.10.3292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Friis R. R. Temperature-sensitive mutants of avian RNA tumor viruses: a review. Curr Top Microbiol Immunol. 1978;79:261–293. doi: 10.1007/978-3-642-66853-1_6. [DOI] [PubMed] [Google Scholar]
  18. Fusco A., Portella G., Di Fiore P. P., Berlingieri M. T., Di Lauro R., Schneider A. B., Vecchio G. A mos oncogene-containing retrovirus, myeloproliferative sarcoma virus, transforms rat thyroid epithelial cells and irreversibly blocks their differentiation pattern. J Virol. 1985 Oct;56(1):284–292. doi: 10.1128/JVI.56.1.284-292.1985. [DOI] [PMC free article] [PubMed] [Google Scholar] [Retracted]
  19. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  20. Hamelin R., Kabat K., Blair D., Arlinghaus R. B. Temperature-sensitive splicing defect of ts110 Moloney murine sarcoma virus is virus encoded. J Virol. 1986 Jan;57(1):301–309. doi: 10.1128/jvi.57.1.301-309.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hannink M., Donoghue D. J. Lysine residue 121 in the proposed ATP-binding site of the v-mos protein is required for transformation. Proc Natl Acad Sci U S A. 1985 Dec;82(23):7894–7898. doi: 10.1073/pnas.82.23.7894. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hwang L. S., Park J., Gilboa E. Role of intron-contained sequences in formation of moloney murine leukemia virus env mRNA. Mol Cell Biol. 1984 Nov;4(11):2289–2297. doi: 10.1128/mcb.4.11.2289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Klein B., Le Bousse C., Fagg B., Smajda-Joffe F., Vehmeyer K., Mori K. J., Jasmin C., Ostertag W. Effects of myeloproliferative sarcoma virus on the pluripotential stem cell and granulocyte precursor cell populations of DBA/2 mice. J Natl Cancer Inst. 1981 May;66(5):935–940. [PubMed] [Google Scholar]
  24. Kloetzer W. S., Maxwell S. A., Arlinghaus R. B. Further characterization of the P85gag-mos -associated protein kinase activity. Virology. 1984 Oct 15;138(1):143–155. doi: 10.1016/0042-6822(84)90154-5. [DOI] [PubMed] [Google Scholar]
  25. Kloetzer W. S., Maxwell S. A., Arlinghaus R. B. P85gag-mos encoded by ts110 Moloney murine sarcoma virus has an associated protein kinase activity. Proc Natl Acad Sci U S A. 1983 Jan;80(2):412–416. doi: 10.1073/pnas.80.2.412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kollek R., Stocking C., Smadja-Joffe F., Ostertag W. Molecular cloning and characterization of a leukemia-inducing myeloproliferative sarcoma virus and two of its temperature-sensitive mutants. J Virol. 1984 Jun;50(3):717–724. doi: 10.1128/jvi.50.3.717-724.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mann R., Mulligan R. C., Baltimore D. Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus. Cell. 1983 May;33(1):153–159. doi: 10.1016/0092-8674(83)90344-6. [DOI] [PubMed] [Google Scholar]
  28. Maxwell S. A., Arlinghaus R. B. Serine kinase activity associated with Maloney murine sarcoma virus-124-encoded p37mos. Virology. 1985 May;143(1):321–333. doi: 10.1016/0042-6822(85)90119-9. [DOI] [PubMed] [Google Scholar]
  29. Maxwell S. A., Arlinghaus R. B. Use of site-specific antipeptide antibodies to perturb the serine kinase catalytic activity of p37mos. J Virol. 1985 Sep;55(3):874–876. doi: 10.1128/jvi.55.3.874-876.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Moelling K., Heimann B., Beimling P., Rapp U. R., Sander T. Serine- and threonine-specific protein kinase activities of purified gag-mil and gag-raf proteins. Nature. 1984 Dec 6;312(5994):558–561. doi: 10.1038/312558a0. [DOI] [PubMed] [Google Scholar]
  31. Nishizawa M., Mayer B. J., Takeya T., Yamamoto T., Toyoshima K., Hanafusa H., Kawai S. Two independent mutations are required for temperature-sensitive cell transformation by a Rous sarcoma virus temperature-sensitive mutant. J Virol. 1985 Dec;56(3):743–749. doi: 10.1128/jvi.56.3.743-749.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Oskarsson M., McClements W. L., Blair D. G., Maizel J. V., Vande Woude G. F. Properties of a normal mouse cell DNA sequence (sarc) homologous to the src sequence of Moloney sarcoma virus. Science. 1980 Mar 14;207(4436):1222–1224. doi: 10.1126/science.6243788. [DOI] [PubMed] [Google Scholar]
  33. Ostertag W., Freshney M., Vehmeyer K., Jasmin C., Rutter G. Action of temperature-sensitive mutants of myeloproliferative sarcoma virus suggests that fibroblast-transforming and hematopoietic transforming viral properties are related. J Virol. 1984 Jan;49(1):253–261. doi: 10.1128/jvi.49.1.253-261.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ostertag W., Seliger B., Kollek R., Stocking C., Bergholz U., Smadja-Joffe F. The myeloproliferative sarcoma virus retains transforming functions after introduction of a dominant selectable marker gene. J Gen Virol. 1986 Jul;67(Pt 7):1361–1371. doi: 10.1099/0022-1317-67-7-1361. [DOI] [PubMed] [Google Scholar]
  35. Ostertag W., Stocking C., Johnson G. R., Kluge N., Kollek R., Franz T., Hess N. Transforming genes and target cells of murine spleen focus-forming viruses. Adv Cancer Res. 1987;48:193–355. doi: 10.1016/s0065-230x(08)60693-4. [DOI] [PubMed] [Google Scholar]
  36. Ostertag W., Vehmeyer K., Fagg B., Pragnell I. B., Paetz W., Le Bousse M. C., Smadja-Joffe F., Klein B., Jasmin C., Eisen H. Myeloproliferative virus, a cloned murine sarcoma virus with spleen focus-forming properties in adult mice. J Virol. 1980 Feb;33(2):573–582. doi: 10.1128/jvi.33.2.573-582.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Papkoff J., Hunter T., Beemon K. In vitro translation of virion RNA from Moloney murine sarcoma virus. Virology. 1980 Feb;101(1):91–103. doi: 10.1016/0042-6822(80)90486-9. [DOI] [PubMed] [Google Scholar]
  38. Papkoff J., Nigg E. A., Hunter T. The transforming protein of Moloney murine sarcoma virus is a soluble cytoplasmic protein. Cell. 1983 May;33(1):161–172. doi: 10.1016/0092-8674(83)90345-8. [DOI] [PubMed] [Google Scholar]
  39. Papkoff J., Verma I. M., Hunter T. Detection of a transforming gene product in cells transformed by Moloney murine sarcoma virus. Cell. 1982 Jun;29(2):417–426. doi: 10.1016/0092-8674(82)90158-1. [DOI] [PubMed] [Google Scholar]
  40. Pragnell I. B., Fusco A., Arbuthnott C., Smadja-Joffe F., Klein B., Jasmin C., Ostertag W. Analysis of the myeloproliferative sarcoma virus genome: limited changes in the prototype lead to altered target cell specificity. J Virol. 1981 Jun;38(3):952–957. doi: 10.1128/jvi.38.3.952-957.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Propst F., Vande Woude G. F. Expression of c-mos proto-oncogene transcripts in mouse tissues. Nature. 1985 Jun 6;315(6019):516–518. doi: 10.1038/315516a0. [DOI] [PubMed] [Google Scholar]
  42. Reddy E. P., Smith M. J., Aaronson S. A. Complete nucleotide sequence and organization of the Moloney murine sarcoma virus genome. Science. 1981 Oct 23;214(4519):445–450. doi: 10.1126/science.6170110. [DOI] [PubMed] [Google Scholar]
  43. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Scolnick E. M., Stephenson J. R., Aaronson S. A. Isolation of temperature-sensitive mutants of murine sarcoma virus. J Virol. 1972 Oct;10(4):653–657. doi: 10.1128/jvi.10.4.653-657.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Seliger B., Kollek R., Stocking C., Franz T., Ostertag W. Viral transfer, transcription, and rescue of a selectable myeloproliferative sarcoma virus in embryonal cell lines: expression of the mos oncogene. Mol Cell Biol. 1986 Jan;6(1):286–293. doi: 10.1128/mcb.6.1.286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Seth A., Vande Woude G. F. Nucleotide sequence and biochemical activities of the Moloney murine sarcoma virus strain HT-1 mos gene. J Virol. 1985 Oct;56(1):144–152. doi: 10.1128/jvi.56.1.144-152.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Stacey A., Arbuthnott C., Kollek R., Coggins L., Ostertag W. Comparison of myeloproliferative sarcoma virus with Moloney murine sarcoma virus variants by nucleotide sequencing and heteroduplex analysis. J Virol. 1984 Jun;50(3):725–732. doi: 10.1128/jvi.50.3.725-732.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Stocking C., Kollek R., Bergholz U., Ostertag W. Long terminal repeat sequences impart hematopoietic transformation properties to the myeloproliferative sarcoma virus. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5746–5750. doi: 10.1073/pnas.82.17.5746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Stocking C., Kollek R., Bergholz U., Ostertag W. Point mutations in the U3 region of the long terminal repeat of Moloney murine leukemia virus determine disease specificity of the myeloproliferative sarcoma virus. Virology. 1986 Aug;153(1):145–149. doi: 10.1016/0042-6822(86)90015-2. [DOI] [PubMed] [Google Scholar]
  50. Topp W. C. Normal rat cell lines deficient in nuclear thymidine kinase. Virology. 1981 Aug;113(1):408–411. doi: 10.1016/0042-6822(81)90168-9. [DOI] [PubMed] [Google Scholar]

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

RESOURCES