Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

Notchic-ER chimeras display hormone-dependent transformation, nuclear accumulation, phosphorylation and CBF1 activation

Oncogene volume 19, pages 3914–3924 (2000)Cite this article

Abstract

Notch genes encode a family of evolutionarily conserved transmembrane receptors that are involved in many distinct cellular processes such as differentiation, proliferation and apoptosis. Notch function has been shown to be required both during development and in adult life. Moreover, several studies on spontaneous human tumors and in experimental models demonstrate that three of the four mammalian Notch genes can act as oncogenes. The mechanism by which Notch proteins induce neoplastic transformation is not known. In order to determine the early signaling events mediated by Notch during cellular transformation we constructed several inducible alleles of Notchic by fusing portions of Nic to the hormone-binding domain of the estrogen receptor. Here we show that Notchic-ER chimeras are conditionally activated by 4-Hydroxytamoxifen (OHT) in a dose-dependent manner. Clonal RKE cell lines expressing Notchic-ER chimeras display hormone-dependent transformation in vitro. Transformation mediated by Notchic-ER is reversible and chronic stimulation is necessary for the maintenance of the transformed phenotype. In response to hormone activation Notchic-ER chimeras become hyperphosphorylated and accumulate in the nucleus of the cell; indicating that both phosphorylation and nuclear localization are required for Notch transforming activity.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  • Ahmad I, Zagouras P and Artavanis-Tsakonas S. . 1995 Mech. Dev. 53: 73–85.

  • Arnold SF, Obourn JD, Jaffe H and Notides AC. . 1995 Mol. Endocrinol. 9: 24–33.

  • Artavanis-Tsakonas S, Matsuno K and Fortini ME. . 1995 Science 268: 225–232.

  • Artavanis-Tsakonas S, Rand MD and Lake RJ. . 1999 Science 284: 770–776.

  • Aster J, Pear W, Hasserjian R, Erba H, Davi F, Luo B, Scott M, Baltimore D and Sklar J. . 1994 Cold Spring Harb. Symp. Quant. Biol. 59: 125–136.

  • Burk O and Klempnauer KH. . 1991 EMBO J. 10: 3713–3719.

  • Capobianco AJ and Gilmore TD. . 1993 Virology 193: 160–170.

  • Capobianco AJ, Zagouras P, Blaumueller CM, Artavanis-Tsakonas S and Bishop JM. . 1997 Mol. Cell. Biol. 17: 6265–6273.

  • Chernova OB, Chernov MV, Ishizaka Y, Agarwal ML and Stark GR. . 1998 Mol. Cell. Biol. 18: 536–545.

  • Daniel B, Rangarajan A, Mukherjee G, Vallikad E and Krishna S. . 1997 J. Gen. Virol. 78: 1095–1101.

  • Dievart A, Beaulieu N and Jolicoeur P. . 1999 Oncogene 18: 5973–5981.

  • Dumont E, Fuchs KP, Bommer G, Christoph B, Kremmer E and Kempkes B. . 2000 Oncogene 19: 556–561.

  • Eccles SA, Modjtahedi H, Box G, Court W, Sandle J and Dean CJ. . 1994 Invasion Metastasis 14: 337–348.

  • Eilers M, Picard D, Yamamoto KR and Bishop JM. . 1989 Nature 340: 66–68.

  • Ellisen LW, Bird J, West DC, Soreng AL, Reynolds TC, Smith SD and Sklar J. . 1991 Cell 66: 649–661.

  • Fehon RG, Johansen K, Rebay I and Artavanis-Tsakonas S. . 1991 J. Cell. Biol. 113: 657–669.

  • Felsher DW and Bishop JM. . 1999 Proc. Natl. Acad. Sci. USA 96: 3940–3944.

  • Fortini ME and Artavanis-Tsakonas S. . 1994 Cell 79: 273–282.

  • Fortini ME, Rebay I, Caron LA and Artavanis-Tsakonas S. . 1993 Nature 365: 555–557.

  • Fuentes-Panana EM and Ling PD. . 1998 J. Virol. 72: 693–700.

  • Gallahan D, Kozak C and Callahan R. . 1987 J. Virol. 61: 218–220.

  • Girard L, Hanna Z, Beaulieu N, Hoemann CD, Simard C, Kozak CA and Jolicoeur P. . 1996 Genes. Dev. 10: 1930–1944.

  • Grignani F, Kinsella T, Mencarelli A, Valtieri M, Riganelli D, Lanfrancone L, Peschle C, Nolan GP and Pelicci PG. . 1998 Cancer Res. 58: 14–19.

  • Heitzler P and Simpson P. . 1991 Cell 64: 1083–1092.

  • Hsieh JJ, Henkel T, Salmon P, Robey E, Peterson MG and Hayward SD. . 1996 Mol. Cell. Biol. 16: 952–959.

  • Jackson P, Baltimore D and Picard D. . 1993 EMBO J. 12: 2809–2819.

  • Jarriault S, Brou C, Logeat F, Schroeter EH, Kopan R and Israel A. . 1995 Nature 377: 355–358.

  • Jeffries S and Capobianco AJ. . 2000 Mol. Cell. Biol. 20: 3928–3941.

  • Jhappan C, Gallahan D, Stahle C, Chu E, Smith GH, Merlino G and Callahan R. . 1992 Genes Dev. 6: 345–355.

  • Kao HY, Ordentlich P, Koyano-Nakagawa N, Tang Z, Downes M, Kintner CR, Evans RM and Kadesch T. . 1998 Genes Dev. 12: 2269–2277.

  • Kidd S, Baylies MK, Gasic GP and Young MW. . 1989 Genes Dev. 3: 1113–1129.

  • Kidd S, Lieber T and Young MW. . 1998 Genes Dev. 12: 3728–3740.

  • Kopan R, Nye JS and Weintraub H. . 1994 Development 120: 2385–2396.

  • Kopan R, Schroeter EH, Weintraub H and Nye JS. . 1996 Proc. Natl. Acad. Sci. USA 93: 1683–1688.

  • Kruse U, Iacovoni JS, Goller ME and Vogt PK. . 1997 Proc. Natl. Acad. Sci. USA 94: 12396–12400.

  • Le Goff P, Montano MM, Schodin DJ and Katzenellenbogen BS. . 1994 J. Biol. Chem. 269: 4458–4466.

  • Lecourtois M and Schweisguth F. . 1998 Curr. Biol. 8: 771–774.

  • Lieber T, Kidd S, Alcamo E, Corbin V and Young MW. . 1993 Genes Dev. 7: 1949–1965.

  • Lofts FJ and Gullick WJ. . 1992 Cancer Treat Res. 61: 161–179.

  • Miele L and Osborne B. . 1999 J. Cell Physiol. 181: 393–409.

  • Milner LA and Bigas A. . 1999 Blood 93: 2431–2448.

  • Nye JS, Kopan R and Axel R. . 1994 Development 120: 2421–2430.

  • Oswald F, Liptay S, Adler G and Schmid RM. . 1998 Mol. Cell. Biol. 18: 2077–2088.

  • Pear WS, Aster JC, Scott ML, Hasserjian RP, Soffer B, Sklar J and Baltimore D. . 1996 J. Exp. Med. 183: 2283–2291.

  • Rebay I, Fehon RG and Artavanis-Tsakonas S. . 1993 Cell 74: 319–329.

  • Rehberger S, Gounari F, DucDodon M, Chlichlia K, Gazzolo L, Schirrmacher V and Khazaie K. . 1997 Exp. Cell. Res. 233: 363–371.

  • Robbins J, Blondel BJ, Gallahan D and Callahan R. . 1992 J. Virol. 66: 2594–2599.

  • Rohn JL, Lauring AS, Linenberger ML and Overbaugh J. . 1996 J. Virol. 70: 8071–8080.

  • Samuels ML, Weber MJ, Bishop JM and McMahon M. . 1993 Mol. Cell. Biol. 13: 6241–6252.

  • Schroeter EH, Kisslinger JA and Kopan R. . 1998 Nature 393: 382–386.

  • Smith GH, Gallahan D, Diella F, Jhappan C, Merlino G and Callahan R. . 1995 Cell Growth Differ. 6: 563–577.

  • Stifani S, Blaumueller CM, Redhead NJ, Hill RE and Artavanis-Tsakonas S. . 1992 Nat. Genet. 2: 343.

  • Struhl G and Adachi A. . 1998 Cell 93: 649–660.

  • Struhl G, Fitzgerald K and Greenwald I. . 1993 Cell 74: 331–345.

  • Superti-Furga G, Bergers G, Picard D and Busslinger M. . 1991 Proc. Natl. Acad. Sci. USA 88: 5114–5118.

  • Tamura K, Taniguchi Y, Minoguchi S, Sakai T, Tun T, Furukawa T and Hongo T. . 1995 Curr. Biol. 5: 1416–1423.

  • Vigo E, Muller H, Prosperini E, Hateboer G, Cartwright P, Moroni MC and Helin K. . 1999 Mol. Cell. Biol. 19: 6379–6395.

  • Zagouras P, Stifani S, Blaumueller CM, Carcangiu ML and Artavanis-Tsakonas S. . 1995 Proc. Natl. Acad. Sci. USA 92: 6414–6418.

Download references

Acknowledgements

We thank members of the Capobianco laboratory for support and technical assistance. We thank David Robbins and his laboratory for insightful comments on our work. We also thank Gary Nolan (Stanford), Paul Ling (Baylor) and Martin MacMahon (UCSF) for kindly providing reagents used in this study. This work was funded in part by grants from the American Cancer Society (RPG LBC-99465 to AJ Capobianco and the National Cancer Institute (ROI CA 83736 to AJ Capobianco).

Author information

Authors and Affiliations

  1. Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati, College of Medicine, Cincinnati, 45267-0524, Ohio, OH, USA

    Chiara Ronchini & Anthony J Capobianco

Authors
  1. Chiara Ronchini
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anthony J Capobianco
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ronchini, C., Capobianco, A. Notchic-ER chimeras display hormone-dependent transformation, nuclear accumulation, phosphorylation and CBF1 activation. Oncogene 19, 3914–3924 (2000). https://doi.org/10.1038/sj.onc.1203719

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1203719

Keywords

This article is cited by

Search

Advanced search

Quick links