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:

Identification of multiple SNT-binding sites on NPM-ALK oncoprotein and their involvement in cell transformation

Abstract

The t(2;5) chromosomal translocation occurs in anaplastic large-cell lymphoma arising from activated T lymphocytes. This genomic rearrangement generates the nucleophosmin (NPM)-anaplastic lymphoma kinase (ALK) oncoprotein that is a chimeric protein consisting of parts of the nuclear protein NPM and ALK receptor protein-tyrosine kinase. We used yeast two-hybrid screening to identify an adaptor protein Suc1-associated neurotrophic factor-induced tyrosine-phosphorylated target (SNT)-2 as a new partner that interacted with the cytoplasmic domain of ALK. Immunoprecipitation assay revealed that SNT-1 and SNT-2 interacted with NPM-ALK and kinase-negative NPM-ALK mutant. Y156, Y567 and a 19-amino-acid sequence (aa 631–649) of NPM-ALK were essential for this interaction. The interaction through Y156 and Y567 was dependent on phosphorylation of these tyrosines, whereas the interaction through the 19-amino-acid sequence was independent of phosphorylation. NPM-ALK mutant protein mutated at these three binding sites showed significantly reduced transforming activity. This transformation-defective NPM-ALK mutant still interacted with signal transducing proteins such as phospholipase C-gamma and phosphatidylinositol 3-kinase, which were previously reported to be relevant to NPM-ALK-dependent tumorigenesis. These observations indicate that the three SNT-binding sites of NPM-ALK are important for its transforming activity. This raises a possibility that SNT family proteins play significant roles in cellular transformation triggered by NPM-ALK, which though remains to be verified.

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

Similar content being viewed by others

References

  • Bai RY, Dieter P, Peschel C, Morris SW, Duyster J . (1998). Nucleophosmin-anaplastic lymphoma kinase of large-cell anaplastic lymphoma is a constitutively active tyrosine kinase that utilizes phospholipase C-gamma to mediate its mitogenicity. Mol Cell Biol 18: 6951–6961.

    Article  CAS  Google Scholar 

  • Bai RY, Ouyang T, Miething C, Morris SW, Peschel C, Duyster J . (2000). Nucleophosmin-anaplastic lymphoma kinase associated with anaplastic large-cell lymphoma activates the phosphatidylinositol 3-kinase/Akt antiapoptotic signaling pathway. Blood 96: 4319–4327.

    CAS  PubMed  Google Scholar 

  • Dhalluin C, Yan K, Plotnikova O, Lee KW, Zeng L, Kuti M et al. (2000). Structural basis of SNT PTB domain interactions with distinct neurotrophic receptors. Mol Cell 6: 921–929.

    Article  CAS  Google Scholar 

  • Fujimoto J, Shiota M, Iwahara T, Seki N, Satoh H, Mori S et al. (1996). Characterization of the transforming activity of p80, a hyperphosphorylated protein in a Ki-1 lymphoma cell line with chromosomal translocation t(2;5). Proc Natl Acad Sci USA 93: 4181–4186.

    Article  CAS  Google Scholar 

  • Gotoh N, Laks S, Nakashima M, Lax I, Schlessinger J . (2004). FRS2 family docking proteins with overlapping roles in activation of MAP kinase have distinct spatial-temporal patterns of expression of their transcripts. FEBS Lett 564: 14–18.

    Article  CAS  Google Scholar 

  • Hadari YR, Kouhara H, Lax I, Schlessinger J . (1998). Binding of Shp2 tyrosine phosphatase to FRS2 is essential for fibroblast growth factor-induced PC12 cell differentiation. Mol Cell Biol 18: 3966–3973.

    Article  CAS  Google Scholar 

  • Herbst H, Dallenbach F, Hummel M, Niedobitek G, Finn T, Young LS et al. (1991). Epstein–Barr virus DNA and latent gene products in Ki-1 (CD30)-positive anaplastic large cell lymphomas. Blood 78: 2666–2673.

    CAS  PubMed  Google Scholar 

  • Huang L, Gotoh N, Zhang S, Shibuya M, Yamamoto T, Tsuchida N . (2004). SNT-2 interacts with ERK2 and negatively regulates ERK2 signaling in response to EGF stimulation. Biochem Biophys Res Commun 324: 1011–1017.

    Article  CAS  Google Scholar 

  • Iwahara T, Fujimoto J, Wen D, Cupples R, Bucay N, Arakawa T et al. (1997). Molecular characterization of ALK, a receptor tyrosine kinase expressed specifically in the nervous system. Oncogene 14: 439–449.

    Article  CAS  Google Scholar 

  • Kao S, Jaiswal RK, Kolch W, Landreth GE . (2001). Identification of the mechanisms regulating the differential activation of the mapk cascade by epidermal growth factor and nerve growth factor in PC12 cells. J Biol Chem 276: 18169–18177.

    Article  CAS  Google Scholar 

  • Kurokawa K, Iwashita T, Murakami H, Hayashi H, Kawai K, Takahashi M . (2001). Identification of SNT/FRS2 docking site on RET receptor tyrosine kinase and its role for signal transduction. Oncogene 20: 1929–1938.

    Article  CAS  Google Scholar 

  • Lax I, Wong A, Lamothe B, Lee A, Frost A, Hawes J et al. (2002). The docking protein FRS2alpha controls a MAP kinase-mediated negative feedback mechanism for signaling by FGF receptors. Mol Cell 10: 709–719.

    Article  CAS  Google Scholar 

  • McDougall K, Kubu C, Verdi JM, Meaken SO . (2001). Developmental expression patterns of the signaling adapters FRS-2 and FRS-3 during early embryogenesis. Mech Dev 103: 145–158.

    Article  CAS  Google Scholar 

  • Meakin SO, MacDonald JI, Gryz EA, Kubu CJ, Verdi JM . (1999). The signaling adapter FRS-2 competes with Shc for binding to the nerve growth factor receptor TrkA. A model for discriminating proliferation and differentiation. J Biol Chem 274: 9861–9870.

    Article  CAS  Google Scholar 

  • Morris SW, Kirstein MN, Valentine MB, Dittmer KG, Shapiro DN, Saltman DL et al. (1994). Fusion of a kinase gene, ALK, to a nucleolar protein gene, NPM, in non-Hodgkin's lymphoma. Science 263: 1281–1284.

    Article  CAS  Google Scholar 

  • Morris SW, Naeve C, Mathew P, James PL, Kirstein MN, Cui X et al. (1997). ALK, the chromosome 2 gene locus altered by the t(2;5) in non-Hodgkin's lymphoma, encodes a novel neural receptor tyrosine kinase that is highly related to leukocyte tyrosine kinase (LTK). Oncogene 14: 2175–2188.

    Article  CAS  Google Scholar 

  • Ong SH, Guy GR, Hadari YR, Laks S, Gotoh N, Schlessinger J et al. (2000). FRS2 proteins recruit intracellular signaling pathways by binding to diverse targets on fibroblast growth factor and nerve growth factor receptors. Mol Cell Biol 20: 979–989.

    Article  CAS  Google Scholar 

  • Shiota M, Fujimoto J, Semba T, Satoh H, Yamamoto T, Mori S . (1994). Hyperphosphorylation of a novel 80 kDa protein-tyrosine kinase similar to Ltk in a human Ki-1 lymphoma cell line, AMS3. Oncogene 9: 1567–1574.

    CAS  Google Scholar 

  • Slupianek A, Nieborowska-Skorska M, Hoser G, Morrione A, Majewski M, Xue L et al. (2001). Role of phosphatidylinositol 3-kinase–Akt pathway in nucleophosmin/anaplastic lymphoma kinase-mediated lymphomagenesis. Cancer Res 61: 2194–2199.

    CAS  PubMed  Google Scholar 

  • Zamo A, Chiarle R, Piva R, Howes J, Fan Y, Chilosi M et al. (2002). Anaplastic lymphoma kinase (ALK) activates Stat3 and protects hematopoietic cells from cell death. Oncogene 21: 1038–1047.

    Article  CAS  Google Scholar 

  • Zhang Q, Raghunath PN, Xue L, Majewski M, Carpentieri DF, Odum N et al. (2002). Multilevel dysregulation of STAT3 activation in anaplastic lymphoma kinase-positive T/null-cell lymphoma. J Immunol 168: 466–474.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Noriko Gotoh for valuable discussion. This study was supported by Grant-in-Aid for Scientific Research on Priority Areas for Cancer from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to T Yamamoto.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chikamori, M., Fujimoto, J., Tokai-Nishizumi, N. et al. Identification of multiple SNT-binding sites on NPM-ALK oncoprotein and their involvement in cell transformation. Oncogene 26, 2950–2954 (2007). https://doi.org/10.1038/sj.onc.1210095

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1210095

Keywords

This article is cited by

Search

Quick links