title: Systematic Identification of Oncogenic EGFR Interaction Partners.
abstract: The epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase (TK) that-once activated upon ligand binding-leads to receptor dimerization, recruitment of protein complexes, and activation of multiple signaling cascades. The EGFR is frequently overexpressed or mutated in various cancers leading to aberrant signaling and tumor growth. Hence, identification of interaction partners that bind to mutated EGFR can help identify novel targets for drug discovery. Here, we used a systematic approach to identify novel proteins that are involved in cancerous EGFR signaling. Using a combination of high-content imaging and a mammalian membrane two-hybrid protein-protein interaction method, we identified eight novel interaction partners of EGFR, of which half strongly interacted with oncogenic, hyperactive EGFR variants. One of these, transforming acidic coiled-coil proteins (TACC) 3, stabilizes EGFR on the cell surface, which results in an increase in downstream signaling via the mitogen-activated protein kinase and AKT pathway. Depletion of TACC3 from cells using small hairpin RNA (shRNA) knockdown or small-molecule targeting reduced mitogenic signaling in non-small cell lung cancer cell lines, suggesting that targeting TACC3 has potential as a new therapeutic approach for non-small cell lung cancer.
authors.affiliation: MRC Laboratory for Molecular Cell Biology, University College London, London, WC1E 6BT, UK.
authors.forename: Julia
authors.initials: J
authors.lastname: Petschnigg
authors.affiliation: Princess Margaret Cancer Center, University Health Network, Toronto, M5G 2M9, Canada.
authors.forename: Max
authors.initials: M
authors.lastname: Kotlyar
authors.affiliation: MRC Laboratory for Molecular Cell Biology, University College London, London, WC1E 6BT, UK.
authors.forename: Louise
authors.initials: L
authors.lastname: Blair
authors.affiliation: Princess Margaret Cancer Center, University Health Network, Toronto, M5G 2M9, Canada; Department of Medical Biophysics, University of Toronto, Toronto, M5G 1L7, Canada; Department of Computer Science, University of Toronto, Toronto, M5S 2E4, Canada; TECHNA Institute for the Advancement of Technology for Health, Toronto, M5G 1L5, Canada.
authors.forename: Igor
authors.initials: I
authors.lastname: Jurisica
authors.affiliation: Donnelly Centre, Departments of Molecular Genetics and Biochemistry, University of Toronto, Toronto, M5S 3E1, Canada; Department of Molecular Genetics, University of Toronto, M5S 1A8, Canada; Department of Biochemistry, University of Toronto, M5S 1A8, Canada.
authors.forename: Igor
authors.initials: I
authors.lastname: Stagljar
authors.affiliation: MRC Laboratory for Molecular Cell Biology, University College London, London, WC1E 6BT, UK. Electronic address: r.ketteler@ucl.ac.uk.
authors.forename: Robin
authors.initials: R
authors.lastname: Ketteler
mesh_terms: D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics; D002470:Cell Survival; D019295:Computational Biology; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D015972:Gene Expression Regulation, Neoplastic; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D054730:Protein Interaction Domains and Motifs; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D008869:Microtubule-Associated Proteins; D034741:RNA, Small Interfering; C120551:TACC3 protein, human; C512478:EGFR protein, human; D066246:ErbB Receptors; D020928:Mitogen-Activated Protein Kinases
keywords: EGFR; non-small cell lung cancer; oncogenic signaling; protein–protein interaction; two-hybrid screening