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
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
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
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.
Baserga R . (2005). The insulin-like growth factor-I receptor as a target for cancer therapy. Expert Opin Ther Targets 9: 753–768.
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.
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.
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.
Huang EJ, Reichardt LF . (2003). Trk receptors: roles in neuronal signal transduction. Annu Rev Biochem 72: 609–642.
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.
Kerppola TK . (2009). Visualization of molecular interactions using bimolecular fluorescence complementation analysis: characteristics of protein fragment complementation. Chem Soc Rev 38: 2876–2886.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Supplementary Information accompanies the paper on the Oncogene website
Rights 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
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/onc.2011.323
Keywords
This article is cited by
-
The emerging roles of MAPK-AMPK in ferroptosis regulatory network
Cell Communication and Signaling (2023)
-
Fusion partners of NTRK3 affect subcellular localization of the fusion kinase and cytomorphology of melanocytes
Modern Pathology (2021)
-
Oncogenic fusion proteins adopt the insulin-like growth factor signaling pathway
Molecular Cancer (2018)
-
NTRK fusion-positive cancers and TRK inhibitor therapy
Nature Reviews Clinical Oncology (2018)