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Serine mutations that abrogate ligand-induced ubiquitination and internalization of the EGF receptor do not affect c-Cbl association with the receptor

Oncogene volume 22, pages 8509–8518 (2003)Cite this article

Abstract

In the present study, we examined EGF-induced internalization, degradation and trafficking of the epidermal growth factor receptor (EGFR) mutated at serines 1046, 1047, 1057 and 1142 located in its cytoplasmic carboxy-terminal region. We found the serine-mutated EGFR to be inhibited in EGF-induced internalization and degradation in NIH3T3 cells. We therefore tested the hypothesis that these mutations affect ligand-induced c-Cbl association with the receptor, leading to inhibited receptor ubiquitination. EGF was unable to induce ubiquitination of the serine-mutated EGFR, yet EGF-induced phosphorylation of the c-Cbl-binding site at tyrosine 1045, and c-Cbl-EGFR association, was unaffected. To compare the relevance of these serine residues with tyrosine 1045 in their regulation of c-Cbl binding and receptor ubiquitination, we analysed an EGFR mutated at tyrosine 1045 (Y1045F). EGF-induced c-Cbl-EGFR binding was partially inhibited, and receptor ubiquitination was abrogated in cells expressing Y1045F-EGFR. In contrast, ligand-induced internalization and degradation of the Y1045F mutant was similar to that of wild-type EGFR. Together, our data indicate that the serine residues and tyrosine 1045 are essential for EGF-induced receptor ubiquitination, but only the serine residues are critical for EGFR internalization and degradation.

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Acknowledgements

We thank Helga Grøsvik for excellent technical assistance, and Dr Brian Druker for the generous provision of the 4G10 antibody. This work was supported by grants from the Research Council of Norway, the Norwegian Cancer Society, and NH&MRC (Canberra).

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Authors and Affiliations

  1. Department of Pathology, University of Western Australia, Queen Elizabeth II Medical Center, Nedlands, 6009, Western Australia, Australia

    Morten P Oksvold, Christine B F Thien & Wallace Y Langdon

  2. Laboratorium for Toxicopathology, Institute of Pathology, University of Oslo, Rikshospitalet University Hospital, Oslo, N-0027, Norway

    Morten P Oksvold, Jannicke Widerberg & Henrik S Huitfeldt

  3. School of Biological Sciences, University of East Anglia, Norwich, NR4 7TJ, UK

    Andrew Chantry

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  1. Morten P Oksvold
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  2. Christine B F Thien
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Correspondence to Morten P Oksvold.

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Oksvold, M., Thien, C., Widerberg, J. et al. Serine mutations that abrogate ligand-induced ubiquitination and internalization of the EGF receptor do not affect c-Cbl association with the receptor. Oncogene 22, 8509–8518 (2003). https://doi.org/10.1038/sj.onc.1207117

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