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Association of ErbB2 Ser1113 phosphorylation with epidermal growth factor receptor co-expression and poor prognosis in human breast cancer

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Abstract

The carboxyterminal domain of the epidermal growth factor receptor (EGFR) – a putative binding site for the ubiquitin ligase Cbl – is the site of serine phosphorylation events which are essential for ligand-dependent EGFR desensitization and degradation. Using a monoclonal antibody (aPS1113) which selectively recognizes the homologous phosphorylated domain in the ErbB2 oncoprotein, we show here that wild-type ErbB2 becomes Ser1113-phosphorylated following treatment of 3T3 cells with growth factors or tyrosine phosphatase inhibitors. In EGFR-overexpressing A431 cells, ligand-inducible aPS1113 immunoreactivity declines more rapidly than other detectable phosphorylation events and is followed by EGFR downregulation. Analysis of 65 ErbB2-expressing primary breast cancers reveals a highly significant relationship between Ser1113 phosphorylation and EGFR overexpression (p < 0.0001) as well as an association with poor prognosis (p = 0.005). We submit that ErbB2 Ser1113 phosphorylation status represents a novel and informative biomarker of cancer cell biology and tumor behavior.

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References

  1. Davis RJ, Czech MP: Platelet-derived growth factor mimics phorbol diester action on epidermal growth factor receptor phosphorylation at threonine-654. Proc Natl Acad Sci USA 82: 4080–4084, 1985

    Google Scholar 

  2. Davis RJ, Czech MP: Tumor-promoting phorbol diesters cause the phosphorylation of epidermal growth factor receptors in normal human fibroblasts at threonine-654, Proc Natl Acad Sci USA 82: 1974-1978, 1985

    Google Scholar 

  3. Lin CR, Chen WS, Lazar CS, Carpenter CD, Gill GN, Evans RM, Rosenfeld MG: Protein kinase C phosphorylation at Thr 654 of the unoccupied EGF receptor and EGF binding regulate functional receptor loss by independent mechanisms. Cell 44: 839–848, 1986

    Google Scholar 

  4. Countaway JL, McQuilkin P, Girones N, Davis RJ: Multisite phosphorylation of the epidermal growth factor receptor: use of site-directed mutagenesis to examine the role of serine/threonine phosphorylation. J Biol Chem 265: 3407–3416, 1990

    Google Scholar 

  5. Countaway JL, Nairn AC, Davis RJ: Mechanism of desensitization of the epidermal growth factor receptor protein-tyrosine kinase. J Biol Chem 267: 1129–1140, 1992

    Google Scholar 

  6. Zhou Q, Hakomori S, Kitamura K, Igarashi Y: GM3 directly inhibits tyrosine phosphorylation and de-N-acetyl-GM3 direcetly enhances serine phosphorylation of epidermal growth factor receptor, independently of receptor-receptor interaction. J Biol Chem 269: 1959–1965, 1994

    Google Scholar 

  7. Hernandez-Sotomayor S, Mumby M, Carpenter G: Okadaic acid-induced hyperphosphorylation of the epidermal growth factor receptor. J Biol Chem 266: 21281–21286, 1991

    Google Scholar 

  8. Massoglia S, Gray A, Dull TJ et al.: Epidermal growth factor receptor cytoplasmic domain mutations trigger ligand-independent transformation. Mol Cell Biol 10: 3048–3055, 1990

    Google Scholar 

  9. Theroux SJ, Taglienti-Sian C, Nair N, Countaway JL, Robinson HL, Davis RJ: Increased oncogenic potential of ErbB is associated with the loss of a COOH-terminal domain serine phosphorylation site. J Biol Chem 267: 7967–7970, 1992

    Google Scholar 

  10. Theroux SJ, Latour DA, Stanley K, Raden DL, Davis RJ: Signal transduction by the epidermal growth factor receptor is attenuated by a COOH-terminal domain serine phosphorylation site. J Biol Chem 267: 16620–16626, 1992

    Google Scholar 

  11. Feinmesser RL, Gray K, Means AR, Chantry A: HER-2/erbB2 is phosphorylated by calmodulin-dependent protein kinase II on a single site in the cytoplasmic tail at threonine-1172. Oncogene 12: 2725–2730, 1996

    Google Scholar 

  12. Epstein RJ, Druker BJ, Roberts TM, Stiles CD: Synthetic phosphopeptide immunogens yield activation-specific antibodies to the c-erbB-2 receptor. Proc Natl Acad Sci USA 89: 10435–10439, 1992

    Google Scholar 

  13. Huang GC, Ouyang X, Epstein RJ: Proxy activation of ErbB2 by heterologous ligands suggests a heterotetrameric mechanism of receptor tyrosine kinase interaction. Biochem J 331: 113–119, 1998

    Google Scholar 

  14. Epstein, R. J. Preferential detection of catalytically inactive c-erbB-2 by antibodies to unphoshorylated peptides mimicking receptor tyrosine autophosphorylation sites. Oncogene 11: 315–323, 1995

    Google Scholar 

  15. Gulliford T, Huang GC, Ouyang X, Epstein RJ: Reduced ability of transforming growth factor-alpha to induce hetero-oligomerization and downregulation of EGFR suggests a mechanism of oncogenic synergy with ErbB2. Oncogene 15: 2219–2223, 1997

    Google Scholar 

  16. Ouyang X, Huang GC, Chantry A, Epstein RJ: Adjacent carboxyterminal tyrosine phosphorylation events identify functionally distinct ErbB2 receptor subsets. Exp Cell Res 241: 467–475, 1998

    Google Scholar 

  17. Ouyang X, Gulliford T, Zhang H, Huang GC, Epstein RJ: Human cancer cells exhibit protein kinase C-dependent c-erbB-2 transmodulation which correlate with phosphatase sensitivity and kinase activity. J Biol Chem 271: 21786–21792, 1996

    Google Scholar 

  18. Schechter AL, Stern DF, Vaidyanathan L et al.: The neu oncogene: An erbB-related gene encoding a 185,000-Mr tumour antigen. Nature 312: 513–518, 1984

    Google Scholar 

  19. Towbin H, Staehelin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354, 1979

    Google Scholar 

  20. Coussens L, Yang-Feng TL, Liao YC, Chen E, Gray A, McGrath J, Seeburg PH, Libermann TA, Schlessinger J, Francke U, Ullrich A: Tyrosine kinase receptor with extensive homology to EGF receptor shares chromosomal location with neu oncogene. Science 230: 1132–1139, 1985

    Google Scholar 

  21. Bargmann CI, Weinberg RA: Increased tyrosine kinase activity associated with the protein encoded by the activated neu oncogene. Proc Natl Acad Sci USA 85: 5394–5398, 1988

    Google Scholar 

  22. Weiner DB, Liu J, Cohen JA, Williams WV, Greene MI: A point mutation in the neu oncogene mimics ligand induction of receptor aggregation. Nature 339: 230–231, 1989

    Google Scholar 

  23. Campos-Gonzalez R, Glenney JR: Immunodetection of the ligand-activated receptor for epidermal growth factor. Growth Factors 4: 305-316, 1991

    Google Scholar 

  24. Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL: Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science 235: 177–182, 1987

    Google Scholar 

  25. Qian X, LeVea CM, Freeman JK, Dougall WC, Greene MI: Heterodimerization of epidermal growth factor receptor and wild-type or kinase-deficient Neu: A mechanism of interreceptor kinase activation and transphosphorylation. Proc Natl Acad Sci USA 91: 1500–1504, 1994

    Google Scholar 

  26. Huang GC, Ouyang X, Chantry A, Epstein RJ: Overexpression of ErbB2 impairs downregulation of epidermal growth factor receptors via a post-transcriptional mechanism. J Cell Biochem 74: 23–30, 1999

    Google Scholar 

  27. Lenferink AE, Kramer RH, van Vugt MJ, Konigswieser M, Di Fiore PP, van Zoelen EJ, van de Poll ML: Superagonistic behaviour of epidermal growth factor/transforming growth factor-alpha chimeras: Correlation with receptor routing after ligand-induced internalization. Biochem J 327: 859–865, 1997

    Google Scholar 

  28. Ouyang X, Gulliford T, Huang G, Epstein RJ: Transforming growth factor-alpha short-circuits downregulation of the epidermal growth factor receptor. J Cell Physiol 179: 52–57, 1999

    Google Scholar 

  29. Lemoine NR, Staddon S, Dickson C, Barnes DM, Gullick WJ: Absence of activating transmembrane mutations in the c-erbB-2 proto-oncogene in human breast cancer. Oncogene 5: 237–239, 1990

    Google Scholar 

  30. Langlois WJ, Sasaoka T, Saltiel AR, Olefsky JM: Negative feedback regulation and desensitization of insulin-and epidermal growth factor stimulated p21ras activation. J Biol Chem 270: 25320–25323, 1995

    Google Scholar 

  31. Kuppuswamy D, Dalton M, Pike LJ: Serine 1002 is a site of in vivo and in vitro phosphorylation of the epidermal growth factor receptor. J Biol Chem 268: 19134–19142, 1993

    Google Scholar 

  32. Thien CB, Langdon WY: EGF receptor binding and transformation by v-cbl is ablated by the introduction of a loss-of-function mutation from the Caenorhabditis elegans sli-1 gene. Oncogene 14: 2239–2249, 1997

    Google Scholar 

  33. Ouyang X, Gulliford T, Doherty A, Huang GC, Epstein RJ: Detection of ErbB2 oversignalling in a majority of breast cancers with phosphorylation state-specific antibodies. Lancet 353: 1591–1592, 1999

    Google Scholar 

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Authors and Affiliations

  1. Department of Metabolic Medicine and Oncology, Imperial College School of Medicine, London, UK

    Xiaomei Ouyang, Gillian Smith & Richard J. Epstein

  2. Department of Oncology, Imperial College School of Medicine, London, UK

    Tim Gulliford & Guocai Huang

  3. Department of Biochemistry, Imperial College School of Medicine, London, UK

    Hongyi Zhang

  4. Division of Medical Sciences, National Cancer Centre, Singapore

    Richard J. Epstein

Authors
  1. Xiaomei Ouyang
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  2. Tim Gulliford
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  3. Hongyi Zhang
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  4. Gillian Smith
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  5. Guocai Huang
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  6. Richard J. Epstein
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Correspondence to Richard J. Epstein.

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Ouyang, X., Gulliford, T., Zhang, H. et al. Association of ErbB2 Ser1113 phosphorylation with epidermal growth factor receptor co-expression and poor prognosis in human breast cancer. Mol Cell Biochem 218, 47–54 (2001). https://doi.org/10.1023/A:1007249004222

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  • DOI: https://doi.org/10.1023/A:1007249004222

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