Abstract
The Shc adaptor is responsible for coupling receptor tyrosine kinases and tyrosine kinase-associated receptors to the Ras/MAP kinase pathway. Shc is believed to be regulated by a change in subcellular localization from the cytosol to the plasma membrane, where it recruits Grb-2/Sos complexes and hence permits juxtaposition of the guanine nucleotide exchange factor Sos to Ras, resulting in GDP/GTP exchange and Ras activation. Shc has been recently shown to inducibly colocalize in detergent-resistant membrane rafts together with the activated TCR and associated signaling molecules. To understand whether Shc localization in membrane rafts is sufficient to regulate Shc function, we constructed a Shc chimera containing the Ras membrane localization motif at the C-terminus. We show that membrane targeted Shc was constitutively localized in the plasma membrane of T-cells, and was mostly compartmentalized in lipid rafts. Membrane targeted Shc was phosphorylated on tyrosine residues and bound Grb-2/Sos in the absence of TCR engagement. Furthermore, expression of membrane targeted Shc resulted in constitutive downstream signaling, including Erk2 activation and enhancement of TCR dependent activation of the TCR responsive transcription factor NF-AT. Hence localization of Shc in membrane rafts is sufficient for Shc to acquire a signaling competent state. Interestingly, a membrane targeted Shc mutant lacking both Grb-2 binding sites was not only incapable of signaling in the absence of TCR triggering, but transdominantly inhibited endogenous Shc, supporting a non redundant role for Shc in the activation of the Ras/MAP kinase pathway in T-cells.
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Abbreviations
- PTK:
-
protein tyrosine kinase
- TCR:
-
T-cell antigen receptor
- MAP:
-
mitogen activated protein
- ITAM:
-
immunoreceptor tyrosine-based activation motif
- PTB:
-
phosphotyrosine binding
- CH:
-
collagen homology
- PMA:
-
phorbol myristate acetate
References
Aronheim A, Engelberg D, Li N, al-Alawi N, Schlessinger J and Karin M . 1994 Cell 78: 949–961
Baldari CT, Heguy A and Telford JL . 1993 J Biol Chem 268: 2693–2698
Baldari CT, Pelicci G, Di Somma MM, Milia E, Giuli S, Pelicci PG and Telford JL . 1995 Oncogene 10: 1141–1147
Baldari CT and Telford JL . 1999 Biol Chem 380: 129–134
Bonfini L, Migliaccio E, Pelicci G, Lanfrancone L and Pelicci PG . 1996 TIBS 21: 257–261
Bordier C . 1981 J Biol Chem 256: 1604–1607
Chen RH, Corbalan-Garcia S and Bar-Sagi D . 1997 EMBO J 16: 1351–1359
Crabtree GR and Clipstone NA . 1994 Annu Rev Biochem 63: 1045–1083
Duplay P, Thome M, Herve F and Acuto O . 1994 J Exp Med 179: 1163–1172
Genot E, Cleverley S, Henning S and Cantrell D . 1996 EMBO J 15: 3923–3933
Gotoh N, Tojo A and Shibuya M . 1996 EMBO J 15: 6197–6204
Graef IA, Holsinger LJ, Diver S, Schreiber SL and Crabtree GL . 1997 EMBO J 16: 5618–5628
Hancock JF, Cadwallader K, Paterson H and Marshall CJ . 1991 EMBO J 10: 4033–4039
Harmer SL and DeFranco AL . 1997 Mol Cell Biol 17: 4087–4095
Holsinger LJ, Spencer DM, Austin DJ, Schreiber SL and Crabtree GR 1995 . Proc Natl Acad Sci USA 92: 9810–9814
Iwashima M, Irving BA, van Oers N, Chan AC and Weiss A . 1994 Science 263: 1136–1139
Koretzky GA . 1997 Immunol Today 8: 401–406
Magee T and Marshall C . 1999 Cell 98: 9–12
Marshall CJ . 1994 Curr Opin Genet Dev 4: 82–8
McCormick F . 1993 Nature 363: 15–16
Milia E, Di Somma MM, Baldoni F, Chiari R, Lanfrancone L, Pelicci PG, Telford JL and Baldari CT . 1996 Oncogene 13: 767–775
Montixi C, Langlet C, Bernard AM, Thimonier J, Dubois C, Wurbel MA, Chauvin JP, Pierres M and He HT . 1998 EMBO J 17: 5334–5348
Neumeister EN, Zhu Y, Richard S, Terhost C, Chan AC and Shaw AS . 1995 Mol Cell Biol 15: 3171–3178
Pacini S, Ulivieri C, Di Somma MM, Isacchi A, Lanfrancone L, Pelicci PG, Telford JL and Baldari CT . 1998 J Biol Chem 273: 20487–20493
Pawson T . 1995 Nature 373: 573–580
Peterson EJ, Clements JL, Fang N and Koretzky GA . 1998 Curr Opin Immunol 10: 337–344
Ravichandran K, Lee KK, Songyang Z, Cantley LC, Burn P and Burakoff SJ . 1993 Science 262: 902–905
Ravichandran K, Lorenz U, Shoelson SE and Burakoff SJ . 1995 Mol Cell Biol 15: 593–600
Ravichandran KS, Zhou MM, Pratt JC, Harlan JA, Walk SF, Fesik SW and Burakoff SJ . 1997 Mol Cell Biol 17: 5540–5549
Rudd CE . 1999 Cell 96: 5–8
Simons K and Ikonen E . 1997 Nature 387: 569–572
Ulivieri C, Pacini S, Bartalini S, Valensin S, Telford JL and Baldari CT . 1999 Eur J Immunol 29: in press
Walk SF, March ME and Ravichandran KS . 1998 Eur J Immunol 28: 2265–2275
Wange RL and Samelson LE . 1996 Immunity 5: 197–205
Weiss A and Littman DR . 1994 Cell 76: 263–274
Wessel D and Flugge UL . 1983 Anal Biochem 138: 141–143
Woodrow MA, Rayter S, Downward J and Cantrell DA 1993a . J Immunol 150: 3853–3861
Xavier R, Brennan T, Li Q, McCormack C and Seed B . 1998 Immunity 8: 723–732
Yamasaki S, Takamatsu M and Iwashima M . 1996 Mol Cell Biol 16: 7151–7160
Zhang W, Sloan-Lancaster J, Kitchen J, Trible RP and Samelson LE . 1998 Cell 92: 83–92
Acknowledgements
The authors wish to acknowledge S Grassini for technical assistance, L Gamberucci for contribution to the artwork, and G Benocci for secretarial assistance. They also indebted to T Pawson for the generous gift of the HA-tagged Shc cDNA and to JL Telford for productive discussions, helpful advise and critical reading of the manuscript. This work was generously supported by the Italian Association for Cancer Research (AIRC) and Telethon (grant n. E.651). The contribution of the MURST (quota 60 and 40%) is also gratefully acknowledged. S Plyte is the recipient of an EMBO long-term fellowship. MB Majolini and S Pacini are the recipients of a fellowship from the Italian Federation for Cancer Research (FIRC).
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Plyte, S., Majolini, M., Pacini, S. et al. Constitutive activation of the Ras/MAP kinase pathway and enhanced TCR signaling by targeting the Shc adaptor to membrane rafts. Oncogene 19, 1529–1537 (2000). https://doi.org/10.1038/sj.onc.1203451
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DOI: https://doi.org/10.1038/sj.onc.1203451
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