Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1989 Sep;8(9):2575–2581. doi: 10.1002/j.1460-2075.1989.tb08396.x

Reversible abrogation of IL-3 dependence by an inducible H-ras oncogene.

E Andrejauskas 1, C Moroni 1
PMCID: PMC401260  PMID: 2555182

Abstract

Immortalized, interleukin-3 (IL-3)-dependent mouse mast cells (PB-3c) were transfected with a human activated c-H-ras gene under the transcriptional control of the mouse mammary tumor virus long terminal repeat. Addition of increasing amounts of dexamethasone resulted in a concentration-dependent increase in expression of the H-ras oncogene. The elevation of p21 ras protein concentrations was paralleled by progressive growth of the transfectants in the absence of exogenous IL-3, leading to complete abrogation of growth-factor requirement at high p21ras levels. The maintenance of the IL-3-independent state required the continuous expression of the H-ras oncogene, since dexamethasone removal was followed by rapid cell death. Expression of the H-ras oncogene induced PB-3c cells to produce IL-3 and granulocyte-macrophage colony-stimulating factor, suggesting that their IL-3-independent proliferation may be due to an autocrine mechanism.

Full text

PDF
2580

Images in this article

2576Image
on p.2576
2576Image
on p.2576
2578Image
on p.2578
2578Image
on p.2578

Selected References

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

  1. Anzano M. A., Roberts A. B., De Larco J. E., Wakefield L. M., Assoian R. K., Roche N. S., Smith J. M., Lazarus J. E., Sporn M. B. Increased secretion of type beta transforming growth factor accompanies viral transformation of cells. Mol Cell Biol. 1985 Jan;5(1):242–247. doi: 10.1128/mcb.5.1.242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ball P. E., Conroy M. C., Heusser C. H., Davis J. M., Conscience J. F. Spontaneous, in vitro, malignant transformation of a basophil/mast cell line. Differentiation. 1983;24(1):74–78. doi: 10.1111/j.1432-0436.1983.tb01305.x. [DOI] [PubMed] [Google Scholar]
  3. Bishop J. M. The molecular genetics of cancer. Science. 1987 Jan 16;235(4786):305–311. doi: 10.1126/science.3541204. [DOI] [PubMed] [Google Scholar]
  4. Bowen-Pope D. F., Vogel A., Ross R. Production of platelet-derived growth factor-like molecules and reduced expression of platelet-derived growth factor receptors accompany transformation by a wide spectrum of agents. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2396–2400. doi: 10.1073/pnas.81.8.2396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cepko C. L., Roberts B. E., Mulligan R. C. Construction and applications of a highly transmissible murine retrovirus shuttle vector. Cell. 1984 Jul;37(3):1053–1062. doi: 10.1016/0092-8674(84)90440-9. [DOI] [PubMed] [Google Scholar]
  6. Cohen J. B., Levinson A. D. A point mutation in the last intron responsible for increased expression and transforming activity of the c-Ha-ras oncogene. Nature. 1988 Jul 14;334(6178):119–124. doi: 10.1038/334119a0. [DOI] [PubMed] [Google Scholar]
  7. Conscience J. F., Fischer F. Growth-factor-independent proliferation in vitro and tumorigenicity in vivo are associated in basophil/mast-cell lines and their somatic hybrids. Differentiation. 1985;28(3):291–295. doi: 10.1111/j.1432-0436.1985.tb00838.x. [DOI] [PubMed] [Google Scholar]
  8. Cook W. D., Metcalf D., Nicola N. A., Burgess A. W., Walker F. Malignant transformation of a growth factor-dependent myeloid cell line by Abelson virus without evidence of an autocrine mechanism. Cell. 1985 Jul;41(3):677–683. doi: 10.1016/s0092-8674(85)80048-9. [DOI] [PubMed] [Google Scholar]
  9. Cory S., Bernard O., Bowtell D., Schrader S., Schrader J. W. Murine c-myc retroviruses alter the growth requirements of myeloid cell lines. Oncogene Res. 1987 Jun;1(1):61–76. [PubMed] [Google Scholar]
  10. Davis J. M., Pennington J. E., Kubler A. M., Conscience J. F. A simple, single-step technique for selecting and cloning hybridomas for the production of monoclonal antibodies. J Immunol Methods. 1982;50(2):161–171. doi: 10.1016/0022-1759(82)90222-8. [DOI] [PubMed] [Google Scholar]
  11. Dexter T. M., Garland J., Scott D., Scolnick E., Metcalf D. Growth of factor-dependent hemopoietic precursor cell lines. J Exp Med. 1980 Oct 1;152(4):1036–1047. doi: 10.1084/jem.152.4.1036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Furth M. E., Davis L. J., Fleurdelys B., Scolnick E. M. Monoclonal antibodies to the p21 products of the transforming gene of Harvey murine sarcoma virus and of the cellular ras gene family. J Virol. 1982 Jul;43(1):294–304. doi: 10.1128/jvi.43.1.294-304.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gibbs J. B., Sigal I. S., Poe M., Scolnick E. M. Intrinsic GTPase activity distinguishes normal and oncogenic ras p21 molecules. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5704–5708. doi: 10.1073/pnas.81.18.5704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Heldin C. H., Westermark B. Growth factors: mechanism of action and relation to oncogenes. Cell. 1984 May;37(1):9–20. doi: 10.1016/0092-8674(84)90296-4. [DOI] [PubMed] [Google Scholar]
  15. Hume C. R., Nocka K. H., Sorrentino V., Lee J. S., Fleissner E. Constitutive c-myc expression enhances the response of murine mast cells to IL-3, but does not eliminate their requirement for growth factors. Oncogene. 1988 Mar;2(3):223–226. [PubMed] [Google Scholar]
  16. Hurley J. B., Simon M. I., Teplow D. B., Robishaw J. D., Gilman A. G. Homologies between signal transducing G proteins and ras gene products. Science. 1984 Nov 16;226(4676):860–862. doi: 10.1126/science.6436980. [DOI] [PubMed] [Google Scholar]
  17. Isfort R. J., Stevens D., May W. S., Ihle J. N. Interleukin 3 binds to a 140-kDa phosphotyrosine-containing cell surface protein. Proc Natl Acad Sci U S A. 1988 Nov;85(21):7982–7986. doi: 10.1073/pnas.85.21.7982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jaggi R., Salmons B., Muellener D., Groner B. The v-mos and H-ras oncogene expression represses glucocorticoid hormone-dependent transcription from the mouse mammary tumor virus LTR. EMBO J. 1986 Oct;5(10):2609–2616. doi: 10.1002/j.1460-2075.1986.tb04541.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kipreos E. T., Wang J. Y. Reversible dependence on growth factor interleukin-3 in myeloid cells expressing temperature sensitive v-abl oncogene. Oncogene Res. 1988 Feb;2(3):277–284. [PubMed] [Google Scholar]
  20. Koyasu S., Tojo A., Miyajima A., Akiyama T., Kasuga M., Urabe A., Schreurs J., Arai K., Takaku F., Yahara I. Interleukin 3-specific tyrosine phosphorylation of a membrane glycoprotein of Mr 150,000 in multi-factor-dependent myeloid cell lines. EMBO J. 1987 Dec 20;6(13):3979–3984. doi: 10.1002/j.1460-2075.1987.tb02740.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Land H., Chen A. C., Morgenstern J. P., Parada L. F., Weinberg R. A. Behavior of myc and ras oncogenes in transformation of rat embryo fibroblasts. Mol Cell Biol. 1986 Jun;6(6):1917–1925. doi: 10.1128/mcb.6.6.1917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lee J. C., Hapel A. J., Ihle J. N. Constitutive production of a unique lymphokine (IL 3) by the WEHI-3 cell line. J Immunol. 1982 Jun;128(6):2393–2398. [PubMed] [Google Scholar]
  23. Mathey-Prevot B., Nabel G., Palacios R., Baltimore D. Abelson virus abrogation of interleukin-3 dependence in a lymphoid cell line. Mol Cell Biol. 1986 Nov;6(11):4133–4135. doi: 10.1128/mcb.6.11.4133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McKay I. A., Marshall C. J., Calés C., Hall A. Transformation and stimulation of DNA synthesis in NIH-3T3 cells are a titratable function of normal p21N-ras expression. EMBO J. 1986 Oct;5(10):2617–2621. doi: 10.1002/j.1460-2075.1986.tb04542.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Metcalf D., Roberts T. M., Cherington V., Dunn A. R. The in vitro behavior of hemopoietic cells transformed by polyoma middle T antigen parallels that of primary human myeloid leukemic cells. EMBO J. 1987 Dec 1;6(12):3703–3709. doi: 10.1002/j.1460-2075.1987.tb02704.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Morla A. O., Schreurs J., Miyajima A., Wang J. Y. Hematopoietic growth factors activate the tyrosine phosphorylation of distinct sets of proteins in interleukin-3-dependent murine cell lines. Mol Cell Biol. 1988 May;8(5):2214–2218. doi: 10.1128/mcb.8.5.2214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nair A. P., Diamantis I. D., Conscience J. F., Kindler V., Hofer P., Moroni C. A v-H-ras-dependent hemopoietic tumor model involving progression from a clonal stage of transformation competence to autocrine interleukin 3 production. Mol Cell Biol. 1989 Mar;9(3):1183–1190. doi: 10.1128/mcb.9.3.1183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Park L. S., Friend D., Gillis S., Urdal D. L. Characterization of the cell surface receptor for a multi-lineage colony-stimulating factor (CSF-2 alpha). J Biol Chem. 1986 Jan 5;261(1):205–210. [PubMed] [Google Scholar]
  30. Pierce J. H., Di Fiore P. P., Aaronson S. A., Potter M., Pumphrey J., Scott A., Ihle J. N. Neoplastic transformation of mast cells by Abelson-MuLV: abrogation of IL-3 dependence by a nonautocrine mechanism. Cell. 1985 Jul;41(3):685–693. doi: 10.1016/s0092-8674(85)80049-0. [DOI] [PubMed] [Google Scholar]
  31. Rapp U. R., Cleveland J. L., Brightman K., Scott A., Ihle J. N. Abrogation of IL-3 and IL-2 dependence by recombinant murine retroviruses expressing v-myc oncogenes. Nature. 1985 Oct 3;317(6036):434–438. doi: 10.1038/317434a0. [DOI] [PubMed] [Google Scholar]
  32. Rein A., Keller J., Schultz A. M., Holmes K. L., Medicus R., Ihle J. N. Infection of immune mast cells by Harvey sarcoma virus: immortalization without loss of requirement for interleukin-3. Mol Cell Biol. 1985 Sep;5(9):2257–2264. doi: 10.1128/mcb.5.9.2257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Reynolds V. L., Lebovitz R. M., Warren S., Hawley T. S., Godwin A. K., Lieberman M. W. Regulation of a metallothionein-rasT24 fusion gene by zinc results in graded alterations in cell morphology and growth. Oncogene. 1987;1(3):323–330. [PubMed] [Google Scholar]
  34. Schuler G. D., Cole M. D. GM-CSF and oncogene mRNA stabilities are independently regulated in trans in a mouse monocytic tumor. Cell. 1988 Dec 23;55(6):1115–1122. doi: 10.1016/0092-8674(88)90256-5. [DOI] [PubMed] [Google Scholar]
  35. Shaw G., Kamen R. A conserved AU sequence from the 3' untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell. 1986 Aug 29;46(5):659–667. doi: 10.1016/0092-8674(86)90341-7. [DOI] [PubMed] [Google Scholar]
  36. Spandidos D. A., Wilkie N. M. Malignant transformation of early passage rodent cells by a single mutated human oncogene. Nature. 1984 Aug 9;310(5977):469–475. doi: 10.1038/310469a0. [DOI] [PubMed] [Google Scholar]
  37. Stocking C., Löliger C., Kawai M., Suciu S., Gough N., Ostertag W. Identification of genes involved in growth autonomy of hematopoietic cells by analysis of factor-independent mutants. Cell. 1988 Jun 17;53(6):869–879. doi: 10.1016/s0092-8674(88)90329-7. [DOI] [PubMed] [Google Scholar]
  38. Sweet R. W., Yokoyama S., Kamata T., Feramisco J. R., Rosenberg M., Gross M. The product of ras is a GTPase and the T24 oncogenic mutant is deficient in this activity. Nature. 1984 Sep 20;311(5983):273–275. doi: 10.1038/311273a0. [DOI] [PubMed] [Google Scholar]
  39. Wasylyk C., Imler J. L., Perez-Mutul J., Wasylyk B. The c-Ha-ras oncogene and a tumor promoter activate the polyoma virus enhancer. Cell. 1987 Feb 13;48(3):525–534. doi: 10.1016/0092-8674(87)90203-0. [DOI] [PubMed] [Google Scholar]
  40. Watson J. D., Eszes M., Overell R., Conlon P., Widmer M., Gillis S. Effect of infection with murine recombinant retroviruses containing the v-src oncogene on interleukin 2- and interleukin 3-dependent growth states. J Immunol. 1987 Jul 1;139(1):123–129. [PubMed] [Google Scholar]
  41. Weinberg R. A. The action of oncogenes in the cytoplasm and nucleus. Science. 1985 Nov 15;230(4727):770–776. doi: 10.1126/science.2997917. [DOI] [PubMed] [Google Scholar]
  42. Wheeler E. F., Askew D., May S., Ihle J. N., Sherr C. J. The v-fms oncogene induces factor-independent growth and transformation of the interleukin-3-dependent myeloid cell line FDC-P1. Mol Cell Biol. 1987 May;7(5):1673–1680. doi: 10.1128/mcb.7.5.1673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. 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 The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES