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.

  • Short Communication
  • Published:

A tripartite complex composed of ETV6-NTRK3, IRS1 and IGF1R is required for ETV6-NTRK3-mediated membrane localization and transformation

Abstract

ETV6-NTRK3 (EN), a chimeric tyrosine kinase generated by t(12;15) translocations, is a dominantly acting oncoprotein in diverse tumor types. We previously showed that insulin-like growth factor 1 receptor (IGF1R) is essential for EN-mediated oncogenesis and that insulin receptor substrate 1 (IRS1) is constitutively tyrosine phosphorylated and bound by EN in transformed cells. Given that IRS1 is also an adapter for IGF1R, we hypothesized that IRS1 might localize EN to IGF1R at the membrane to activate phosphatidylinositol 3-kinase (PI3K)-Akt, which is critical for EN oncogenesis. In this study, we examined EN/IRS1/IGF1R complexes in detail. We find that both IRS1 and kinase active IGF1R are required for EN transformation, that tyrosine phosphorylated IRS1 is present in high molecular weight complexes with EN and IGF1R, and that EN colocalizes with IGF1R at the plasma membrane. Both IGF1R kinase activity and an intact cytoplasmic Y950 residue, the IRS1-docking site of IGF1R, are required, confirming the importance of the IGF1R/IRS1 interaction for EN oncogenesis. The dual specificity IGF1R and insulin receptor (INSR) inhibitor, BMS-536924, blocks EN transformation activity, cell survival and its interaction with IRS proteins, and induces a striking shift of EN proteins to smaller sized molecular complexes. We conclude that a tripartite complex of EN, IRS1 and IGF1R localizes EN to the membrane and that this is essential for EN-mediated transformation. These findings provide an explanation for the observed IGF1R dependency of EN transformation. Blocking IGF1R kinase activity may, therefore, provide a tractable therapeutic strategy for the many tumor types driven by the EN oncoprotein.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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

Figure 1
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

References

  • Argani P, Fritsch M, Kadkol SS, Schuster A, Beckwith JB, Perlman EJ . (2000). Detection of the ETV6-NTRK3 chimeric RNA of infantile fibrosarcoma/cellular congenital mesoblastic nephroma in paraffin-embedded tissue: application to challenging pediatric renal stromal tumors. Mod Pathol 13: 29–36.

    Article  CAS  Google Scholar 

  • Baserga R . (2005). The insulin-like growth factor-I receptor as a target for cancer therapy. Expert Opin Ther Targets 9: 753–768.

    Article  CAS  Google Scholar 

  • Craparo A, O'Neill TJ, Gustafson TA . (1995). Non-SH2 domains within insulin receptor substrate-1 and SHC mediate their phosphotyrosine-dependent interaction with the NPEY motif of the insulin-like growth factor I receptor. J Biol Chem 270: 15639–15643.

    Article  CAS  Google Scholar 

  • Eguchi M, Eguchi-Ishimae M, Tojo A, Morishita K, Suzuki K, Sato Y et al. (1999). Fusion of ETV6 to neurotrophin-3 receptor TRKC in acute myeloid leukemia with t(12;15)(p13;q25). Blood 93: 1355–1363.

    CAS  Google Scholar 

  • Forghieri F, Morselli M, Potenza L, Maccaferri M, Pedrazzi L, Paolini A et al. (2011). Chronic eosinophilic leukaemia with Etv6-Ntrk3 fusion transcript in an elderly patient affected with pancreatic carcinoma. Eur J Haematol 86: 352–355.

    Article  CAS  Google Scholar 

  • Huang EJ, Reichardt LF . (2003). Trk receptors: roles in neuronal signal transduction. Annu Rev Biochem 72: 609–642.

    Article  CAS  Google Scholar 

  • Kazakov DV, Hantschke M, Vanecek T, Kacerovska D, Michal M . (2010). Mammary-type secretory carcinoma of the skin. Am J Surg Pathol 34: 1226–1227; author reply 8.

    Article  Google Scholar 

  • Kerppola TK . (2009). Visualization of molecular interactions using bimolecular fluorescence complementation analysis: characteristics of protein fragment complementation. Chem Soc Rev 38: 2876–2886.

    Article  CAS  Google Scholar 

  • Knezevich SR, Garnett MJ, Pysher TJ, Beckwith JB, Grundy PE, Sorensen PH . (1998). ETV6-NTRK3 gene fusions and trisomy 11 establish a histogenetic link between mesoblastic nephroma and congenital fibrosarcoma. Cancer Res 58: 5046–5048.

    CAS  PubMed  Google Scholar 

  • Knezevich SR, McFadden DE, Tao W, Lim JF, Sorensen PH . (1998). A novel ETV6-NTRK3 gene fusion in congenital fibrosarcoma. Nat Genet 18: 184–187.

    Article  CAS  Google Scholar 

  • Lannon CL, Martin MJ, Tognon CE, Jin W, Kim SJ, Sorensen PH . (2004). A highly conserved NTRK3 C-terminal sequence in the ETV6-NTRK3 oncoprotein binds the phosphotyrosine binding domain of insulin receptor substrate-1: an essential interaction for transformation. J Biol Chem 279: 6225–6234.

    Article  CAS  Google Scholar 

  • Lannon CL, Sorensen PH . (2005). ETV6-NTRK3: a chimeric protein tyrosine kinase with transformation activity in multiple cell lineages. Semin Cancer Biol 15: 215–223.

    Article  CAS  Google Scholar 

  • Martin MJ, Melnyk N, Pollard M, Bowden M, Leong H, Podor TJ et al. (2006). The insulin-like growth factor I receptor is required for Akt activation and suppression of anoikis in cells transformed by the ETV6-NTRK3 chimeric tyrosine kinase. Mol Cell Biol 26: 1754–1769.

    Article  CAS  Google Scholar 

  • Morrison KB, Tognon C, Garnett MJ, Deal C, Sorensen PHB . (2002). ETV6-NTRK3 transformation requires insulin-like growth factor 1 receptor signaling and is associated with constitutive IRS-1 tyrosine phosphorylation. Oncogene 21: 5684–5695.

    Article  CAS  Google Scholar 

  • Myers Jr MG, Sun XJ, Cheatham B, Jachna BR, Glasheen EM, Backer JM et al. (1993). IRS-1 is a common element in insulin and insulin-like growth factor-I signaling to the phosphatidylinositol 3′-kinase. Endocrinology 132: 1421–1430.

    Article  CAS  Google Scholar 

  • Skalova A, Vanecek T, Sima R, Laco J, Weinreb I, Perez-Ordonez B et al. (2010). Mammary analogue secretory carcinoma of salivary glands, containing the ETV6-NTRK3 fusion gene: a hitherto undescribed salivary gland tumor entity. Am J Surg Pathol 34: 599–608.

    PubMed  Google Scholar 

  • Sun H, Baserga R . (2004). Deletion of the pleckstrin and phosphotyrosine binding domains of insulin receptor substrate-2 does not impair its ability to regulate cell proliferation in myeloid cells. Endocrinology 145: 5332–5343.

    Article  CAS  Google Scholar 

  • Tognon C, Garnett M, Kenward E, Kay R, Morrison K, Sorensen PH . (2001). The chimeric protein tyrosine kinase ETV6-NTRK3 requires both Ras-Erk1/2 and PI3-kinase-Akt signaling for fibroblast transformation. Cancer Res 61: 8909–8916.

    CAS  PubMed  Google Scholar 

  • Tognon C, Knezevich SR, Huntsman D, Roskelley CD, Melnyk N, Mathers JA et al. (2002). Expression of the ETV6-NTRK3 gene fusion as a primary event in human secretory breast carcinoma. Cancer Cell 2: 367–376.

    Article  CAS  Google Scholar 

  • Tognon CE, Mackereth C, Somasiri AM, McIntosh LP, Sorensen PHB . (2004). Mutations in the SAM domain of the ETV6-NTRK3 chimeric tyrosine kinase block polymerization and transformation activity. Moll Cell Biol 24: 4636–4650.

    Article  CAS  Google Scholar 

  • Tognon CE, Somasiri AM, Evdokimova VE, Trigo G, Uy EE, Melnyk N et al. (2011). ETV6-NTRK3-mediated breast epithelial cell transformation is blocked by targeting the IGF1R signaling pathway. Cancer Res 71: 1060–1070.

    Article  CAS  Google Scholar 

  • Wai DH, Knezevich SR, Lucas T, Jansen B, Kay RJ, Sorensen PHB . (2000). The ETV6-NTRK3 gene fusion encodes a chimeric protein tyrosine kinase that transforms NIH3T3 cells. Oncogene 19: 906–915.

    Article  CAS  Google Scholar 

  • Wittman M, Carboni J, Attar R, Balasubramanian B, Balimane P, Brassil P et al. (2005). Discovery of a (1H-benzoimidazol-2-yl)-1H-pyridin-2-one (BMS-536924) inhibitor of insulin-like growth factor I receptor kinase with in vivo antitumor activity. J Med Chem 48: 5639–5643.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the following grants: a ReThink Breast Cancer Career Development Award to CET, a Canadian Institutes of Health Research (CIHR) Graduate Studentship to MJM, a CIHR grant to PHBS, and funds from the British Columbia Cancer Foundation through donations to PHBS from Team Finn and other generous riders in the Ride to Conquer Cancer. We thank Adrian Wan and Darren Saunders for assistance with the BiFC cloning.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to P H B Sorensen.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies the paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tognon, C., Martin, M., Moradian, A. et al. A tripartite complex composed of ETV6-NTRK3, IRS1 and IGF1R is required for ETV6-NTRK3-mediated membrane localization and transformation. Oncogene 31, 1334–1340 (2012). https://doi.org/10.1038/onc.2011.323

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2011.323

Keywords

This article is cited by

Search

Quick links