Abstract
The TrkA NGF receptor extracellular region contains three leucine repeats flanked by cysteine clusters and two immunoglobulin-like domains that are required for specific ligand binding. Deletion of the immunoglobulin-like domains abolishes NGF binding and causes ligand independent activation of the receptor. Here we report a specific mutation that increases the binding affinity of the TrkA receptor for NGF. A change of proline 203 to alanine (P203A) in the linker region between the leucine repeats and the first Ig-like domain increased NGF binding by decreasing the ligand rate of dissociation. This mutated receptor was appropriately expressed on the cell surface and promoted ligand-independent neurite outgrowth in PC12nnr5 cells. The mutant receptor was capable of spontaneous dimerization and was constitutively phosphorylated in the absence of ligand. Moreover, expression of TrkA-P203A receptor in fibroblasts induced DNA synthesis and transformation and generated tumours in nude mice. These data suggest that domains outside of the immunoglobulin-like structure contribute to ligand binding and constitutive activation of Trk receptors.
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References
Arevalo JC, Conde B, Hempstead BL, Chao MV, Martin-Zanca D and Perez P. . 2000 Mol. Cell. Biol. 20: 5908–5916.
Barbacid M. . 1995 Curr. Opin. Cell Biol. 7: 148–155.
Blechman JM, Lev S, Barg J, Eisenstein M, Vaks B, Vogel Z, Givol D and Yarden Y. . 1995 Cell 80: 103–113.
Chao MV. . 2000 J. Neurosci. Res. 59: 353–355.
Coulier F, Kumar R, Ernst M, Klein R, Martin-Zanca D and Barbacid M. . 1990 Mol. Cell. Biol. 10: 4202–4210.
Cunnigham ME and Greene LA. . 1998 EMBO J. 17: 7282–7293.
Eng C and Mulligan LM. . 1997 Hum. Mutat. 9: 97–109.
Ezoe K, Holmes SA, Ho L, Bennett CP, Bolognia JL, Brueton L, Burn J, Falabella R, Gatto EM, Issii N, et al. . 1995 Am. J. Hum. Gen. 56: 58–66.
Greco A, Miranda C, Pagliardini S, Fusetti L, Bongarzone I and Pierotti MA. . 1997 Genes Chromosomes Cancer 19: 112–123.
Greco A, Fusetti L, Miranda C, Villa R, Zanotti S, Pagliardini S and Pierotti MA. . 1998 Oncogene 16: 809–816.
Green SH, Rydel RE, Connolly JL and Greene LA. . 1986 J. Cell Biol. 102: 830–843.
Holden PH, Asopa V, Robertson AGS, Clarke AR, Tyler S, Bennett GS, Brain SD, Wilcock GK, Allen SJ, Smith SKF and Dawbarn D. . 1997 Nature Biotechnol. 15: 668–672.
Kaplan DR and Miller FD. . 2000 Curr. Opin. Neurobiol. 10: 381–391.
Lewin GR and Barde YA. . 1996 Ann. Rev. Neurosci. 19: 289–317.
MacDonald JIS and Meakin SO. . 1996 Mol. Cell. Neurosci. 7: 371–390.
Muenke M, Schell U, Hehr A, Robin NH, Losken HW, Schinzel A, Pulleyn LJ, Rutland P, Reardon W, Malcolm S and Winter RM. . 1994 Nature Genet. 8: 269–274.
Naski MC and Ornitz DM. . 1998 Front. Biosci. 3: 781–794.
Neilson KM and Friesel R. . 1996 J. Biol. Chem. 271: 25049–25057.
Ninkina N, Grashchuck M, Buchman VL and Davies AM. . 1997 J. Biol. Chem. 272: 13019–13025.
Omura T, Heldin CH and Ostman . 1997 J. Biol. Chem. 272: 12676–12682.
Perez P, Coll PM, Hempstead BL, Martin-Zanca D and Chao MV. . 1995 Mol. Cell. Neurosci. 6: 97–105.
Plotnikov AN, Hubbard SR, Schlessinger J and Mohammadi M. . 2000 Cell 101: 413–424.
Plotnikov AN, Schlessinger J, Hubbard SR and Mohammadi M. . 1999 Cell 98: 641–650.
Porter AC and Vaillancourt RR. . 1998 Oncogene 16: 1343–1352.
Robertson SC, Tynan JA and Donoghue DJ. . 2000 Trends Genet. 16: 265–271.
Schneider R and Schweiger M. . 1991 Oncogene 6: 1807–1811.
Siegel PM and Muller WJ. . 1996 Proc. Natl. Acad. Sci. USA 93: 8878–8883.
Smith DK and Xue H. . 1997 J. Mol. Biol. 274: 530–545.
Stauber DJ, DiGabriele AD and Hendrickson WA. . 2000 Proc. Natl. Acad. Sci. USA 97: 49–54.
Urfer R, Tsoulofas P, O'Conell L, Shelton DL, Parada LF and Presta LG. . 1995 EMBO J. 14: 2795–2805.
Van Daalen Wetters T, Hawkins SA, Roussel MF and Sherr CJ. . 1992 EMBO J. 11: 551–557.
Wiesmann C, Ultsch MH, Bass SH and de Vos AM. . 1999 Nature 401: 184–188.
Wilkie AOM, Slaney SF, Oldrigde M, Poole MD, Ashworth GJ, Hockley AD, Hayward RD, David DJ, Pulleyn L and Rutland P. . 1995 Nature Genet. 9: 165–172.
Windish JM, Marksteiner R, Lang ME, Auer B and Schneider R. . 1995a Biochemistry 34: 11256–11263.
Windish JM, Marksteiner R and Schneider R. . 1995b J. Cell Biol. 270: 28133–28138.
Acknowledgements
This work was supported by grants from the Fundacion Ramon Areces and the European Union Program BIO4-CT96-0285. JC Arevalo was a recipient of fellowships from those grants. Grant support for MV Chao and BL Hempstead were from the NIH.
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Arevalo, J., Conde, B., Hempstead, B. et al. A novel mutation within the extracellular domain of TrkA causes constitutive receptor activation. Oncogene 20, 1229–1234 (2001). https://doi.org/10.1038/sj.onc.1204215
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DOI: https://doi.org/10.1038/sj.onc.1204215
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