Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1987 Oct;6(10):2923–2929. doi: 10.1002/j.1460-2075.1987.tb02596.x

Oestradiol stimulates tyrosine phosphorylation and hormone binding activity of its own receptor in a cell-free system.

F Auricchio 1, A Migliaccio 1, M Di Domenico 1, E Nola 1
PMCID: PMC553727  PMID: 3691476

Abstract

Recent experiments have shown that calf uterus oestrogen receptor exists in a tyrosine-phosphorylated hormone binding form and in non-phosphorylated, non-hormone binding form. We report here that physiological concentrations of oestradiol in complex with the receptor stimulate the calf uterus receptor kinase that converts the non-hormone binding receptor into hormone binding receptor through phosphorylation of the receptor on tyrosine. The activity of this enzyme has been followed by reactivation of hormone binding sites and phosphorylation on tyrosine of calf uterus phosphatase-inactivated receptor. Phosphorylation of the receptor has been demonstrated by interaction of kinase 32P-phosphorylated proteins with anti-receptor antibody followed either by sucrose gradient centrifugation or SDS-PAGE of the immunoprecipitated proteins. Hormone stimulation of the kinase is inhibited by receptor occupancy of the anti-oestrogen tamoxifen. Oestradiol-receptor complex increases the affinity of the kinase for the dephosphorylated receptor. Findings of this report are consistent with the observation that several protein tyrosine kinases that are associated with peptide hormone receptors are stimulated by the binding of the hormone to the receptor. This is the first report on the activation of a tyrosine kinase by a steroid hormone. The finding that hormones can regulate their own receptor binding activity through a tyrosine kinase is also new.

Full text

PDF
2923

Images in this article

2924Fig. 2.
on p.2924
2926Fig. 4.
on p.2926
2927Fig. 5.
on p.2927

Selected References

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

  1. Auricchio F., Migliaccio A., Castoria G. Dephosphorylation of oestradiol nuclear receptor in vitro. A hypothesis on the mechanism of action of non-steroidal anti-oestrogens. Biochem J. 1981 Sep 15;198(3):699–702. doi: 10.1042/bj1980699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Auricchio F., Migliaccio A., Castoria G., Lastoria S., Rotondi A. Evidence that in vivo estradiol receptor translocated into nuclei is dephosphorylated and released into cytoplasm. Biochem Biophys Res Commun. 1982 May 14;106(1):149–157. doi: 10.1016/0006-291x(82)92070-8. [DOI] [PubMed] [Google Scholar]
  3. Auricchio F., Migliaccio A., Castoria G., Lastoria S., Schiavone E. ATP-dependent enzyme activating hormone binding of estradiol receptor. Biochem Biophys Res Commun. 1981 Aug 31;101(4):1171–1178. doi: 10.1016/0006-291x(81)91571-0. [DOI] [PubMed] [Google Scholar]
  4. Auricchio F., Migliaccio A., Castoria G., Rotondi A., Di Domenico M. Calmodulin-stimulated estradiol receptor-tyrosine kinase. Methods Enzymol. 1987;139:731–744. doi: 10.1016/0076-6879(87)39123-2. [DOI] [PubMed] [Google Scholar]
  5. Auricchio F., Migliaccio A., Castoria G., Rotondi A., Lastoria S. Direct evidence of in vitro phosphorylation-dephosphorylation of the estradiol-17 beta receptor. Role of Ca2+-calmodulin in the activation of hormone binding sites. J Steroid Biochem. 1984 Jan;20(1):31–35. doi: 10.1016/0022-4731(84)90185-7. [DOI] [PubMed] [Google Scholar]
  6. Auricchio F., Migliaccio A. In vitro inactivation of oestrogen receptor by nuclei: prevention by phosphatase inhibitors. FEBS Lett. 1980 Aug 11;117(1):224–226. doi: 10.1016/0014-5793(80)80950-1. [DOI] [PubMed] [Google Scholar]
  7. Auricchio F., Migliaccio A., Rotondi A. Inactivation of oestrogen receptor in vitro by nuclear dephosphorylation. Biochem J. 1981 Feb 15;194(2):569–574. doi: 10.1042/bj1940569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Baldi A., Boyle D. M., Wittliff J. L. Estrogen receptor is associated with protein and phospholipid kinase activities. Biochem Biophys Res Commun. 1986 Mar 13;135(2):597–606. doi: 10.1016/0006-291x(86)90035-5. [DOI] [PubMed] [Google Scholar]
  9. Bishop J. M. Viral oncogenes. Cell. 1985 Aug;42(1):23–38. doi: 10.1016/s0092-8674(85)80098-2. [DOI] [PubMed] [Google Scholar]
  10. Carpenter G., King L., Jr, Cohen S. Epidermal growth factor stimulates phosphorylation in membrane preparations in vitro. Nature. 1978 Nov 23;276(5686):409–410. doi: 10.1038/276409a0. [DOI] [PubMed] [Google Scholar]
  11. Cuatrecasas P. Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J Biol Chem. 1970 Jun;245(12):3059–3065. [PubMed] [Google Scholar]
  12. Ek B., Heldin C. H. Characterization of a tyrosine-specific kinase activity in human fibroblast membranes stimulated by platelet-derived growth factor. J Biol Chem. 1982 Sep 10;257(17):10486–10492. [PubMed] [Google Scholar]
  13. Green S., Walter P., Kumar V., Krust A., Bornert J. M., Argos P., Chambon P. Human oestrogen receptor cDNA: sequence, expression and homology to v-erb-A. Nature. 1986 Mar 13;320(6058):134–139. doi: 10.1038/320134a0. [DOI] [PubMed] [Google Scholar]
  14. Hunter T., Cooper J. A. Epidermal growth factor induces rapid tyrosine phosphorylation of proteins in A431 human tumor cells. Cell. 1981 Jun;24(3):741–752. doi: 10.1016/0092-8674(81)90100-8. [DOI] [PubMed] [Google Scholar]
  15. Hunter T., Cooper J. A. Protein-tyrosine kinases. Annu Rev Biochem. 1985;54:897–930. doi: 10.1146/annurev.bi.54.070185.004341. [DOI] [PubMed] [Google Scholar]
  16. Jacobs S., Kull F. C., Jr, Earp H. S., Svoboda M. E., Van Wyk J. J., Cuatrecasas P. Somatomedin-C stimulates the phosphorylation of the beta-subunit of its own receptor. J Biol Chem. 1983 Aug 25;258(16):9581–9584. [PubMed] [Google Scholar]
  17. Jordan V. C., Koerner S. Tamoxifen (ICI 46,474) and the human carcinoma 8S oestrogen receptor. Eur J Cancer. 1975 Mar;11(3):205–206. doi: 10.1016/0014-2964(75)90119-x. [DOI] [PubMed] [Google Scholar]
  18. Kasuga M., Karlsson F. A., Kahn C. R. Insulin stimulates the phosphorylation of the 95,000-dalton subunit of its own receptor. Science. 1982 Jan 8;215(4529):185–187. doi: 10.1126/science.7031900. [DOI] [PubMed] [Google Scholar]
  19. Katzenellenbogen J. A., Carlson K. E., Heiman D. F., Robertson D. W., Wei L. L., Katzenellenbogen B. S. Efficient and highly selective covalent labeling of the estrogen receptor with [3H]tamoxifen aziridine. J Biol Chem. 1983 Mar 25;258(6):3487–3495. [PubMed] [Google Scholar]
  20. Migliaccio A., Lastoria S., Moncharmont B., Rotondi A., Auricchio F. Phosphorylation of calf uterus 17 beta-estradiol receptor by endogenous Ca2+-stimulated kinase activating the hormone binding of the receptor. Biochem Biophys Res Commun. 1982 Dec 15;109(3):1002–1010. doi: 10.1016/0006-291x(82)92039-3. [DOI] [PubMed] [Google Scholar]
  21. Migliaccio A., Rotondi A., Auricchio F. Calmodulin-stimulated phosphorylation of 17 beta-estradiol receptor on tyrosine. Proc Natl Acad Sci U S A. 1984 Oct;81(19):5921–5925. doi: 10.1073/pnas.81.19.5921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Migliaccio A., Rotondi A., Auricchio F. Estradiol receptor: phosphorylation on tyrosine in uterus and interaction with anti-phosphotyrosine antibody. EMBO J. 1986 Nov;5(11):2867–2872. doi: 10.1002/j.1460-2075.1986.tb04581.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Moncharmont B., Su J. L., Parikh I. Monoclonal antibodies against estrogen receptor: interaction with different molecular forms and functions of the receptor. Biochemistry. 1982 Dec 21;21(26):6916–6921. doi: 10.1021/bi00269a046. [DOI] [PubMed] [Google Scholar]
  24. Puca G. A., Medici N., Molinari A. M., Moncharmont B., Nola E., Sica V. Estrogen receptor of calf uterus: an easy and fast purification procedure. J Steroid Biochem. 1980 Jan;12:105–113. doi: 10.1016/0022-4731(80)90259-9. [DOI] [PubMed] [Google Scholar]
  25. Ushiro H., Cohen S. Identification of phosphotyrosine as a product of epidermal growth factor-activated protein kinase in A-431 cell membranes. J Biol Chem. 1980 Sep 25;255(18):8363–8365. [PubMed] [Google Scholar]
  26. Van Oosbree T. R., Kim U. H., Mueller G. C. Affinity chromatography of estrogen receptors on diethylstilbestrol-agarose. Anal Biochem. 1984 Feb;136(2):321–327. doi: 10.1016/0003-2697(84)90224-0. [DOI] [PubMed] [Google Scholar]
  27. Walter P., Green S., Greene G., Krust A., Bornert J. M., Jeltsch J. M., Staub A., Jensen E., Scrace G., Waterfield M. Cloning of the human estrogen receptor cDNA. Proc Natl Acad Sci U S A. 1985 Dec;82(23):7889–7893. doi: 10.1073/pnas.82.23.7889. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES