Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1993 Jul;12(7):2809–2819. doi: 10.1002/j.1460-2075.1993.tb05942.x

Hormone-conditional transformation by fusion proteins of c-Abl and its transforming variants.

P Jackson 1, D Baltimore 1, D Picard 1
PMCID: PMC413531  PMID: 8334997

Abstract

Fusion of the hormone binding domain (HBD) of steroid receptors to transcription factors renders them hormone-dependent. We show here that an SH3-deleted, oncogenic variant of the Abl tyrosine kinase becomes hormone-dependent for transformation by fusion to the estrogen receptor (ER) HBD, extending the phenomenon to tyrosine kinases. Surprisingly, fusion of the HBD to the normal, non-transforming c-Abl (IV) protein activated transforming activity in a hormone-dependent fashion. In the presence of hormone, the c-Abl:ER fusion protein was transforming, cytoplasmic and tyrosine phosphorylated, whereas it was non-transforming, nuclear and hypophosphorylated without hormone. We have examined the kinetics of activation of the c-Abl:ER protein and found that protein synthesis is required both for kinase activation and for redistribution of the c-Abl:ER protein from the nucleus to the cytoplasm. We suggest that the activation of c-Abl could be due to HBD-mediated dimerization and/or to the ability to overexpress conditionally the normally toxic c-Abl protein. This novel approach may be applicable to a wide variety of proteins, particularly when activating mutations or physiological inducers are unknown or when the protein is toxic to cells.

Full text

PDF
2809

Images in this article

2810Image
on p.2810
2811Image
on p.2811
2813Image
on p.2813
2814Image
on p.2814
2815Image
on p.2815
2816Image
on p.2816

Selected References

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

  1. Anderson S. M., Rands E., Fincham V. J., Wyke J. A. Isolation of a murine retrovirus with a temperature-sensitive src gene. Virology. 1987 Sep;160(1):263–267. doi: 10.1016/0042-6822(87)90070-5. [DOI] [PubMed] [Google Scholar]
  2. Bolen J. B., Veillette A. A function for the lck proto-oncogene. Trends Biochem Sci. 1989 Oct;14(10):404–407. doi: 10.1016/0968-0004(89)90288-0. [DOI] [PubMed] [Google Scholar]
  3. Burk O., Klempnauer K. H. Estrogen-dependent alterations in differentiation state of myeloid cells caused by a v-myb/estrogen receptor fusion protein. EMBO J. 1991 Dec;10(12):3713–3719. doi: 10.1002/j.1460-2075.1991.tb04939.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Campbell M. L., Li W., Arlinghaus R. B. P210 BCR-ABL is complexed to P160 BCR and ph-P53 proteins in K562 cells. Oncogene. 1990 May;5(5):773–776. [PubMed] [Google Scholar]
  5. Cicchetti P., Mayer B. J., Thiel G., Baltimore D. Identification of a protein that binds to the SH3 region of Abl and is similar to Bcr and GAP-rho. Science. 1992 Aug 7;257(5071):803–806. doi: 10.1126/science.1379745. [DOI] [PubMed] [Google Scholar]
  6. Daley G. Q., Van Etten R. A., Jackson P. K., Bernards A., Baltimore D. Nonmyristoylated Abl proteins transform a factor-dependent hematopoietic cell line. Mol Cell Biol. 1992 Apr;12(4):1864–1871. doi: 10.1128/mcb.12.4.1864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dhut S., Chaplin T., Young B. D. Normal c-abl gene protein--a nuclear component. Oncogene. 1991 Aug;6(8):1459–1464. [PubMed] [Google Scholar]
  8. Eilers M., Picard D., Yamamoto K. R., Bishop J. M. Chimaeras of myc oncoprotein and steroid receptors cause hormone-dependent transformation of cells. Nature. 1989 Jul 6;340(6228):66–68. doi: 10.1038/340066a0. [DOI] [PubMed] [Google Scholar]
  9. Engelman A., Rosenberg N. Isolation of temperature-sensitive Abelson virus mutants by site-directed mutagenesis. Proc Natl Acad Sci U S A. 1987 Nov;84(22):8021–8025. doi: 10.1073/pnas.84.22.8021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fawell S. E., Lees J. A., White R., Parker M. G. Characterization and colocalization of steroid binding and dimerization activities in the mouse estrogen receptor. Cell. 1990 Mar 23;60(6):953–962. doi: 10.1016/0092-8674(90)90343-d. [DOI] [PubMed] [Google Scholar]
  11. Franz W. M., Berger P., Wang J. Y. Deletion of an N-terminal regulatory domain of the c-abl tyrosine kinase activates its oncogenic potential. EMBO J. 1989 Jan;8(1):137–147. doi: 10.1002/j.1460-2075.1989.tb03358.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Goff S. P., Tabin C. J., Wang J. Y., Weinberg R., Baltimore D. Transfection of fibroblasts by cloned Abelson murine leukemia virus DNA and recovery of transmissible virus by recombination with helper virus. J Virol. 1982 Jan;41(1):271–285. doi: 10.1128/jvi.41.1.271-285.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Guan J. L., Trevithick J. E., Hynes R. O. Fibronectin/integrin interaction induces tyrosine phosphorylation of a 120-kDa protein. Cell Regul. 1991 Nov;2(11):951–964. doi: 10.1091/mbc.2.11.951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hatakeyama M., Kono T., Kobayashi N., Kawahara A., Levin S. D., Perlmutter R. M., Taniguchi T. Interaction of the IL-2 receptor with the src-family kinase p56lck: identification of novel intermolecular association. Science. 1991 Jun 14;252(5012):1523–1528. doi: 10.1126/science.2047859. [DOI] [PubMed] [Google Scholar]
  15. Jackson P., Baltimore D. N-terminal mutations activate the leukemogenic potential of the myristoylated form of c-abl. EMBO J. 1989 Feb;8(2):449–456. doi: 10.1002/j.1460-2075.1989.tb03397.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kipreos E. T., Lee G. J., Wang J. Y. Isolation of temperature-sensitive tyrosine kinase mutants of v-abl oncogene by screening with antibodies for phosphotyrosine. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1345–1349. doi: 10.1073/pnas.84.5.1345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kipreos E. T., Wang J. Y. Cell cycle-regulated binding of c-Abl tyrosine kinase to DNA. Science. 1992 Apr 17;256(5055):382–385. doi: 10.1126/science.256.5055.382. [DOI] [PubMed] [Google Scholar]
  18. Kumar V., Chambon P. The estrogen receptor binds tightly to its responsive element as a ligand-induced homodimer. Cell. 1988 Oct 7;55(1):145–156. doi: 10.1016/0092-8674(88)90017-7. [DOI] [PubMed] [Google Scholar]
  19. Kumar V., Green S., Staub A., Chambon P. Localisation of the oestradiol-binding and putative DNA-binding domains of the human oestrogen receptor. EMBO J. 1986 Sep;5(9):2231–2236. doi: 10.1002/j.1460-2075.1986.tb04489.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Li W. J., Kloetzer W. S., Arlinghaus R. B. A novel 53 kDa protein complexed with P210bcr-abl in human chronic myelogenous leukemia cells. Oncogene. 1988 Jun;2(6):559–566. [PubMed] [Google Scholar]
  21. Lugo T. G., Pendergast A. M., Muller A. J., Witte O. N. Tyrosine kinase activity and transformation potency of bcr-abl oncogene products. Science. 1990 Mar 2;247(4946):1079–1082. doi: 10.1126/science.2408149. [DOI] [PubMed] [Google Scholar]
  22. Mayer B. J., Jackson P. K., Baltimore D. The noncatalytic src homology region 2 segment of abl tyrosine kinase binds to tyrosine-phosphorylated cellular proteins with high affinity. Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):627–631. doi: 10.1073/pnas.88.2.627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mayer B. J., Jackson P. K., Van Etten R. A., Baltimore D. Point mutations in the abl SH2 domain coordinately impair phosphotyrosine binding in vitro and transforming activity in vivo. Mol Cell Biol. 1992 Feb;12(2):609–618. doi: 10.1128/mcb.12.2.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Murlas C., Nadel J. A., Roberts J. M. The muscarinic receptors of airway smooth muscle: their characterization in vitro. J Appl Physiol Respir Environ Exerc Physiol. 1982 Apr;52(4):1084–1091. doi: 10.1152/jappl.1982.52.4.1084. [DOI] [PubMed] [Google Scholar]
  25. Pendergast A. M., Muller A. J., Havlik M. H., Clark R., McCormick F., Witte O. N. Evidence for regulation of the human ABL tyrosine kinase by a cellular inhibitor. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5927–5931. doi: 10.1073/pnas.88.13.5927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Picard D., Kumar V., Chambon P., Yamamoto K. R. Signal transduction by steroid hormones: nuclear localization is differentially regulated in estrogen and glucocorticoid receptors. Cell Regul. 1990 Feb;1(3):291–299. doi: 10.1091/mbc.1.3.291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Picard D., Salser S. J., Yamamoto K. R. A movable and regulable inactivation function within the steroid binding domain of the glucocorticoid receptor. Cell. 1988 Sep 23;54(7):1073–1080. doi: 10.1016/0092-8674(88)90122-5. [DOI] [PubMed] [Google Scholar]
  28. Prywes R., Foulkes J. G., Baltimore D. The minimum transforming region of v-abl is the segment encoding protein-tyrosine kinase. J Virol. 1985 Apr;54(1):114–122. doi: 10.1128/jvi.54.1.114-122.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schiff-Maker L., Burns M. C., Konopka J. B., Clark S., Witte O. N., Rosenberg N. Monoclonal antibodies specific for v-abl- and c-abl-encoded molecules. J Virol. 1986 Mar;57(3):1182–1186. doi: 10.1128/jvi.57.3.1182-1186.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Seedorf K., Felder S., Millauer B., Schlessinger J., Ullrich A. Analysis of platelet-derived growth factor receptor domain function using a novel chimeric receptor approach. J Biol Chem. 1991 Jul 5;266(19):12424–12431. [PubMed] [Google Scholar]
  31. Superti-Furga G., Bergers G., Picard D., Busslinger M. Hormone-dependent transcriptional regulation and cellular transformation by Fos-steroid receptor fusion proteins. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5114–5118. doi: 10.1073/pnas.88.12.5114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tapley P., Horwitz A., Buck C., Duggan K., Rohrschneider L. Integrins isolated from Rous sarcoma virus-transformed chicken embryo fibroblasts. Oncogene. 1989 Mar;4(3):325–333. [PubMed] [Google Scholar]
  33. Taylor C. M., Blanchard B., Zava D. T. A simple method to determine whole cell uptake of radiolabelled oestrogen and progesterone and their subcellular localization in breast cancer cell lines in monolayer culture. J Steroid Biochem. 1984 May;20(5):1083–1088. doi: 10.1016/0022-4731(84)90347-9. [DOI] [PubMed] [Google Scholar]
  34. Tora L., Mullick A., Metzger D., Ponglikitmongkol M., Park I., Chambon P. The cloned human oestrogen receptor contains a mutation which alters its hormone binding properties. EMBO J. 1989 Jul;8(7):1981–1986. doi: 10.1002/j.1460-2075.1989.tb03604.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ullrich A., Schlessinger J. Signal transduction by receptors with tyrosine kinase activity. Cell. 1990 Apr 20;61(2):203–212. doi: 10.1016/0092-8674(90)90801-k. [DOI] [PubMed] [Google Scholar]
  36. Umek R. M., Friedman A. D., McKnight S. L. CCAAT-enhancer binding protein: a component of a differentiation switch. Science. 1991 Jan 18;251(4991):288–292. doi: 10.1126/science.1987644. [DOI] [PubMed] [Google Scholar]
  37. Van Etten R. A., Jackson P., Baltimore D. The mouse type IV c-abl gene product is a nuclear protein, and activation of transforming ability is associated with cytoplasmic localization. Cell. 1989 Aug 25;58(4):669–678. doi: 10.1016/0092-8674(89)90102-5. [DOI] [PubMed] [Google Scholar]
  38. Varticovski L., Daley G. Q., Jackson P., Baltimore D., Cantley L. C. Activation of phosphatidylinositol 3-kinase in cells expressing abl oncogene variants. Mol Cell Biol. 1991 Feb;11(2):1107–1113. doi: 10.1128/mcb.11.2.1107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wang J. Y. Negative regulation of c-abl tyrosine kinase by its variable N-terminal amino acids. Oncogene Res. 1988;3(3):293–298. [PubMed] [Google Scholar]
  40. Witte O. N., Goff S., Rosenberg N., Baltimore D. A transformation-defective mutant of Abelson murine leukemia virus lacks protein kinase activity. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4993–4997. doi: 10.1073/pnas.77.8.4993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Yamamoto K. R., Godowski P. J., Picard D. Ligand-regulated nonspecific inactivation of receptor function: a versatile mechanism for signal transduction. Cold Spring Harb Symp Quant Biol. 1988;53(Pt 2):803–811. doi: 10.1101/sqb.1988.053.01.091. [DOI] [PubMed] [Google Scholar]
  42. Yan H., Schlessinger J., Chao M. V. Chimeric NGF-EGF receptors define domains responsible for neuronal differentiation. Science. 1991 Apr 26;252(5005):561–563. doi: 10.1126/science.1850551. [DOI] [PubMed] [Google Scholar]
  43. Yarden Y., Ullrich A. Growth factor receptor tyrosine kinases. Annu Rev Biochem. 1988;57:443–478. doi: 10.1146/annurev.bi.57.070188.002303. [DOI] [PubMed] [Google Scholar]
  44. Yoshinaka Y., Katoh I., Luftig R. B. Murine retrovirus Pr65gag forms a 130K dimer in the absence of disulfide reducing agents. Virology. 1984 Jul 30;136(2):274–281. doi: 10.1016/0042-6822(84)90164-8. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES