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Acidic substitution of the activation loop tyrosines in TrkA supports nerve growth factor-dependent, but not nerve growth factor-independent, differentiation and cell cycle arrest in the human neuroblastoma cell line, SY5Y

Oncogene volume 22, pages 8774–8785 (2003)Cite this article

Abstract

TrkA is the receptor tyrosine kinase (RTK) for nerve growth factor (NGF) and stimulates NGF-dependent cell survival and differentiation in primary neurons and also differentiation of neuroblastomas and apoptosis of medulloblastomas. We have previously shown that aspartic acid and glutamic acid substitution (AspGlu and GluAsp) of the activation loop tyrosines in TrkA (Tyr683 and Tyr684) supports NGF-independent neuritogenesis and cell survival in PC12 cell-derived nnr5 cells. In this study, the AspGlu and GluAsp mutant Trks have been analysed for their ability to support NGF-independent and NGF-dependent neuritogenesis, proliferation and cell signalling in the human neuroblastoma cell line, SY5Y. We find that the AspGlu and GluAsp mutant Trks support NGF-dependent, but not NGF-independent, autophosphorylation, neuritogenic responses and/or inhibit cell cycle progression. The NGF-dependent neuritogenic responses are lower for the mutant Trks (approximately 30–60% for AspGlu and 50–60% for GluAsp), relative to wild-type TrkA. While both the AspGlu and GluAsp mutant Trks support NGF-dependent transient phosphorylation of Shc, PLCγ-1, AKT, FRS2, SH2B as well as prolonged MAP kinase activation, the GluAsp mutant induces stronger NGF-dependent tyrosine phosphorylation of FRS2 and SH2B, as well as a stronger reduction in bromodeoxyuridine (BrdU) incorporation. Collectively, these data suggest that neither absolute levels of receptor autophosphorylation, high levels of TrkA expression nor the activation of a specific signalling pathway is dominant and absolutely essential for neuritogenesis and cell cycle arrest of SY5Y cells.

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References

  • Alessi DR, Andjelkovic M, Caudwell B, Cron P, Morrice N, Cohen P and Hemmings BA . (1996). EMBO J., 15, 6541–6551.

  • Bhattacharyya A, Watson FL, Bradlee TA, Pomeroy SL, Stiles CD and Segal RA . (1997). J. Neurosci., 17, 7007–7016.

  • Biedler JL, Helson L and Spengler BA . (1973). Cancer Res., 33, 2643–2652.

  • Brodeur GM, Seeger RC, Schwab M, Varmus HE and Bishop JM . (1984). Science, 224, 1121–1124.

  • Eggert A, Ikegaki N, Liu X, Chou TT, Lee VM, Trojanowski JQ and Brodeur GM . (2000). Oncogene, 19, 2043–2051.

  • Ehlers M, Kaplan DR, Price D and Koliatsos V . (1995). J. Cell Biol., 130, 149–156.

  • Evan GI, Lewis GK, Ramsay G and Bishop JM . (1985). Mol. Cell. Biol., 5, 3610–3616.

  • Graham FL, Smiley J, Russell WC and Nairn R . (1977). J. Gen. Virol., 36, 59–74.

  • Grimes ML, Beattie E and Mobley WC . (1997). Proc. Natl. Acad. Sci. USA, 94, 9909–9914.

  • Grimes M, Zhou J, Beattie E, Yuen E, Hall D, Valletta J, Topp K, LaVail J, Bunnett N and Mobley W . (1996). J. Neurosci., 16, 7950–7964.

  • Gryz EA and Meakin SO . (2000). Oncogene, 19, 417–430.

  • He TC, Zhou S, da Costa LT, Yu J, Kinzler KW and Vogelstein B . (1998). Proc. Natl. Acad. Sci. USA, 95, 2509–2514.

  • Howe CL, Valletta JS, Rusnak AS and Mobley WC . (2001). Neuron, 32, 801–814.

  • Kogner P, Barbany G, Dominici C, Castello MA, Raschella G and Persson H . (1993). Cancer Res., 53, 2044–2050.

  • Kouhara H, Hadari YR, Spivak-Kroizman T, Schilling J, Bar-Sagi D, Lax I and Schlessinger J . (1997). Cell, 89, 693–702.

  • Lavenius E, Gestblom C, Johansson I, Nanberg E and Pahlman S . (1995). Cell Growth Differ., 6, 727–736.

  • MacDonald JIS, Gryz EA, Kubu CJ, Verdi JM and Meakin SO . (2000). J. Biol. Chem., 24, 18225–18233.

  • Marshall CJ . (1995). Cell, 80, 179–185.

  • Matsushima H and Bogenmann E . (1993). Mol. Cell. Biol., 13, 7447–7456.

  • Meakin SO and MacDonald JI . (1998). J. Neurochem., 71, 1875–1888.

  • Meakin SO, MacDonald JI, Gryz EA, Kubu CJ and Verdi JM . (1999). J. Biol. Chem., 274, 9861–9870.

  • Morooka T and Nishida E . (1998). J. Biol. Chem., 273, 24285–24288.

  • Nakagawara A, Arima M, Azar CG, Scavarda NJ and Brodeur GM . (1992). Cancer Res., 52, 1364–1368.

  • Nakagawara A, Arima-Nakagawara M, Scavarda NJ, Azar CG, Cantor AB and Brodeur GM . (1993). N. Engl. J. Med., 328, 847–854.

  • Nakagawara A, Azar CG, Scavarda NJ and Brodeur GM . (1994). Mol. Cell. Biol., 14, 759–767.

  • Obermeier A, Bradshaw RA, Seedorf K, Choidas A, Schlessinger J and Ullrich A . (1994). EMBO J., 13, 1585–1590.

  • Poluha W, Poluha DK and Ross AH . (1995). Oncogene, 10, 185–189.

  • Qian X, Riccio A, Zhang Y and Ginty DD . (1998). Neuron, 21, 1017–1029.

  • Rabin SJ, Cleghon V and Kaplan DR . (1993). Mol. Cell. Biol., 13, 2203–2213.

  • Ryden M, Sehgal R, Dominici C, Schilling FH, Ibanez CF and Kogner P . (1996). Br. J. Cancer, 74, 773–779.

  • Seeger RC, Brodeur GM, Sather H, Dalton A, Siegel SE, Wong KY and Hammond D . (1985). N. Engl. J. Med., 313, 1111–1116.

  • Sonnenfeld KH and Ishii DN . (1982). J. Neurosci. Res., 8, 375–391.

  • Stephens RM, Loeb DM, Copeland TD, Pawson T, Greene LA and Kaplan DR . (1994). Neuron, 12, 691–705.

  • Strathdee CA, McLeod MR and Hall JR . (1999). Gene, 229, 21–29.

  • Wright JH, Drueckes P, Bartoe J, Zhao Z, Shen SH and Krebs EG . (1997). Mol. Biol. Cell, 8, 1575–1585.

  • Wu C, Lai CF and Mobley WC . (2001). J. Neurosci., 21, 5406–5416.

  • Yamashiro DJ, Nakagawara A, Ikegaki N, Liu XG and Brodeur GM . (1996). Oncogene, 12, 37–41.

  • Yamashita H, Avraham S, Jiang S, Dikic I and Avraham H . (1999). J. Biol. Chem., 274, 15059–15065.

  • Yasui H, Katoh H, Yamaguchi Y, Aoki J, Fujita H, Mori K and Negishi M . (2001). J. Biol. Chem., 276, 15298–15305.

  • York RD, Yao H, Dillon T, Ellig CL, Eckert SP, McCleskey EW and Stork PJ . (1998). Nature, 392, 622–626.

  • Zhang YZ, Moheban DB, Conway BR, Bhattacharyya A and Segal RA . (2000). J. Neurosci., 20, 5671–5678.

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Acknowledgements

We acknowledge J McGlade (Hospital for Sick Children, Toronto), D Ginty (The John Hopkins University School of Medicine, Baltimore, MD) and S Pelech (Kineteck Biotech. Corp.) for their generous gifts of antibodies, as well as Kara Manto and Chunhui Li for technical assistance. This research was supported by an operating grant from the Cancer Research Society Inc. to SOM. EAG was supported with a studentship from the Cancer Research Society Inc. SOM is a scholar of the Canadian Institutes of Health Research.

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  1. Ela A Gryz

    Present address: The Hospital for Sick Children, 555 University Ave., Toronto, Ontario, Canada, M5G 1X8

Authors and Affiliations

  1. Laboratory of Neural Signalling, Cell Biology Group, The Robarts Research Institute, 100 Perth Drive, London, N6A 5K8, Ontario, Canada

    Ela A Gryz & Susan O Meakin

  2. Graduate Program in Neuroscience, The University of Western Ontario, London, N6A 5C1, Ontario, Canada

    Ela A Gryz & Susan O Meakin

  3. Department of Biochemistry, The University of Western Ontario, London, N6A 5C1, Ontario, Canada

    Susan O Meakin

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Correspondence to Susan O Meakin.

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Gryz, E., Meakin, S. Acidic substitution of the activation loop tyrosines in TrkA supports nerve growth factor-dependent, but not nerve growth factor-independent, differentiation and cell cycle arrest in the human neuroblastoma cell line, SY5Y. Oncogene 22, 8774–8785 (2003). https://doi.org/10.1038/sj.onc.1206890

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