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Interaction of oestrogen receptor with the regulatory subunit of phosphatidylinositol-3-OH kinase

Nature volume 407, pages 538541 (28 September 2000) | Download Citation

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Abstract

Oestrogen produces diverse biological effects through binding to the oestrogen receptor (ER)1. The ER is a steroid hormone nuclear receptor, which, when bound to oestrogen, modulates the transcriptional activity of target genes2. Controversy exists, however, concerning whether ER has a role outside the nucleus3, particularly in mediating the cardiovascular protective effects of oestrogen4. Here we show that the ER isoform, ERα, binds in a ligand-dependent manner to the p85α regulatory subunit of phosphatidylinositol-3-OH kinase (PI(3)K). Stimulation with oestrogen increases ERα-associated PI(3)K activity, leading to the activation of protein kinase B/Akt and endothelial nitric oxide synthase (eNOS). Recruitment and activation of PI(3)K by ligand-bound ERα are independent of gene transcription, do not involve phosphotyrosine adapter molecules or src-homology domains of p85α, and extend to other steroid hormone receptors. Mice treated with oestrogen show increased eNOS activity and decreased vascular leukocyte accumulation after ischaemia and reperfusion injury. This vascular protective effect of oestrogen was abolished in the presence of PI(3)K or eNOS inhibitors. Our findings define a physiologically important non-nuclear oestrogen-signalling pathway involving the direct interaction of ERα with PI(3)K.

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References

  1. 1.

    et al. Human oestrogen receptor cDNA: sequence, expression and homology to v-erb-A. Nature 320, 134– 139 (1986).

  2. 2.

    et al. Functional domains of the human estrogen receptor. Cell 51, 941–951 ( 1987).

  3. 3.

    & Specific binding sites for oestrogen at the outer surfaces of isolated endometrial cells. Nature 265, 69–72 (1977).

  4. 4.

    Novel aspects of estrogen action. J. Soc. Gynecol. Investig. 7, S8–9 (2000).

  5. 5.

    et al. cDNA cloning of a novel 85 kd protein that has SH2 domains and regulates binding of PI3-kinase to the PDGF beta-receptor. Cell 65, 75–82 ( 1991).

  6. 6.

    , , & Activation of phosphatidylinositol 3-kinase by insulin. Proc. Natl Acad. Sci. USA 87, 1411–1415 (1990).

  7. 7.

    , , & Nitric oxide production contributes to the angiogenic properties of vascular endothelial growth factor in human endothelial cells. J. Clin. Invest. 100, 3131–3139 ( 1997).

  8. 8.

    et al. Purification and characterization of phosphoinositide 3-kinase from rat liver. J. Biol. Chem. 265, 19704–19711 (1990).

  9. 9.

    , , , & PDGF-dependent tyrosine phosphorylation stimulates production of novel polyphosphoinositides in intact cells. Cell 57, 167–175 ( 1989).

  10. 10.

    & The role of phosphoinositide 3-kinase lipid products in cell function. J. Biol. Chem. 274, 8347–8350 (1999).

  11. 11.

    et al. Protein kinase B kinases that mediate phosphatidylinositol 3,4,5- trisphosphate-dependent activation of protein kinase B. Science 279, 710–714 ( 1998).

  12. 12.

    et al. Phosphoinositide-3-OH kinase-dependent regulation of glycogen synthase kinase 3 and protein kinase B/AKT by the integrin-linked kinase. Proc. Natl Acad. Sci. USA 95, 11211– 11216 (1998).

  13. 13.

    et al. Mechanism of activation of protein kinase B by insulin and IGF-1. EMBO J. 15, 6541– 6551 (1996).

  14. 14.

    , , & Expression of a constitutively active Akt Ser/Thr kinase in 3T3-L1 adipocytes stimulates glucose uptake and glucose transporter 4 translocation. J. Biol. Chem. 271, 31372–31378 (1996).

  15. 15.

    , , , & Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B. Nature 378, 785–789 (1995).

  16. 16.

    et al. Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation. Nature 399, 601–605 (1999).

  17. 17.

    et al. Regulation of endothelium-derived nitric oxide production by the protein kinase Akt. Nature 399, 597 –601 (1999).

  18. 18.

    , & PI(3)K: downstream AKTion blocks apoptosis. Cell 88, 435–437 ( 1997).

  19. 19.

    et al. Estrogen receptor alpha mediates the nongenomic activation of endothelial nitric oxide synthase by estrogen. J. Clin. Invest. 103, 401–406 ( 1999).

  20. 20.

    et al. Impaired B cell development and proliferation in absence of phosphoinositide 3-kinase p85α. Science 283, 393–397 (1999).

  21. 21.

    & Insulin-stimulated production of nitric oxide is inhibited by wortmannin. Direct measurement in vascular endothelial cells. J. Clin. Invest. 98, 894– 898 (1996).

  22. 22.

    et al. Distinct phosphotyrosines on a growth factor receptor bind to specific molecules that mediate different signaling pathways. Cell 69, 413–423 ( 1992).

  23. 23.

    , , & Phosphoinositide 3-kinase binds constitutively to α/β-tubulin and binds to gamma-tubulin in response to insulin. J. Biol. Chem. 270, 25985–25991 (1995).

  24. 24.

    et al. Reduced levels of hsp90 compromise steroid receptor action in vivo. Nature 348, 166– 168 (1990).

  25. 25.

    et al. Dynamic activation of endothelial nitric oxide synthase by Hsp90. Nature 392, 821– 824 (1998).

  26. 26.

    & Protein kinase B (c-Akt) in phosphatidylinositol-3-OH kinase signal transduction. Nature 376, 599–602 ( 1995).

  27. 27.

    & Akt mediates cytoprotection of endothelial cells by vascular endothelial growth factor in an anchorage-dependent manner. J. Biol. Chem. 274, 16349– 16354 (1999).

  28. 28.

    , & An absolute requirement for P-selectin in ischemia/reperfusion-induced leukocyte recruitment in cremaster muscle. Microcirculation 5, 281–287 (1998).

  29. 29.

    , & Nitric oxide: an endogenous modulator of leukocyte adhesion. Proc. Natl Acad. Sci. USA 88, 4651–4655 (1991).

  30. 30.

    et al. Cell-surface estrogen receptors mediate calcium-dependent nitric oxide release in human endothelia. Circulation 101, 1594–1597 (2000).

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Acknowledgements

We thank T. Uchida, A. J. Prorock and K. L. Thomas for technical assistance; M. White for providing IRS-1/2 antibodies; M. Brown for ERα antibody; D. Fruman and L. Cantley for murine p85α-/- fibroblasts, GST–p85α and sub-domains; M. Kasuga for wild-type and dominant-negative p85α cDNAs; and K. Walsh for adenovirus Akt mutants. This work was supported by grants from the National Institutes of Health, the Mary Horrigan Connors Center for Women's Health, the American Heart Association and the Scuola Superiore di Studi e di Perfezionamento “S. Anna”.

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Affiliations

  1. *Cardiovascular and

    • Tommaso Simoncini
    •  & James K. Liao
  2. §Genetics Divisions, Department of Medicine, Brigham & Women's Hospital and Harvard Medical School, 221 Longwood Avenue, Boston, Massachusetts 02115, USA

    • William W. Chin
  3. †Department of Biomedical Engineering, University of Virginia Health Sciences Center, Charlottesville , Virginia 22908, USA

    • Ali Hafezi-Moghadam
    •  & Klaus Ley
  4. ‡Howard Hughes Medical Institute, Joslin Diabetes Center, Boston, Massachusetts 02215, USA

    • Derek P. Brazil

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Correspondence to James K. Liao.

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https://doi.org/10.1038/35035131

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