title: Mutation specific functions of EGFR result in a mutation-specific downstream pathway activation.
abstract: BACKGROUND
  Epidermal growth factor receptor (EGFR) is frequently mutated in various types of cancer. Although all oncogenic mutations are considered activating, different tumour types have different mutation spectra. It is possible that functional differences underlie this tumour-type specific mutation spectrum.
  
  
  METHODS
  We have determined whether specific mutations in EGFR (EGFR, EGFRvIII and EGFR-L858R) have differences in binding partners, differences in downstream pathway activation (gene expression and phosphoproteins), and have functional consequences on cellular growth and migration.
  
  
  RESULTS
  Using biotin pulldown and subsequent mass spectrometry we were able to detect mutation specific binding partners for EGFR. Differential binding was confirmed using a proximity ligation assay and/or Western Blot for the dedicator of cytokinesis 4 (DOCK4), UDP-glucose glycoprotein glucosyltransferase 1 (UGGT1), MYC binding protein 2 (MYCBP2) and Smoothelin (SMTN). We also demonstrate that each mutation induces the expression of a specific set of genes, and that each mutation is associated with specific phosphorylation patterns. Finally, we demonstrate using stably expressing cell lines that EGFRvIII and EGFL858R display reduced growth and migration compared to EGFR wildtype expressing cells.
  
  
  CONCLUSIONS
  Our results indicate that there are distinct functional differences between different EGFR mutations. The functional differences between different mutations argue for the development of mutation specific targeted therapies.
authors.affiliation: Dept. of Neurology, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Lale
authors.initials: L
authors.lastname: Erdem-Eraslan
authors.affiliation: Dept. of Neurology, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Ya
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Dept. of Neurology, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Nanne K
authors.initials: NK
authors.lastname: Kloosterhof
authors.affiliation: Dept. of Pathology, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Yassar
authors.initials: Y
authors.lastname: Atlasi
authors.affiliation: Dept. of Proteomics Center, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Jeroen
authors.initials: J
authors.lastname: Demmers
authors.affiliation: Dept. of Pathology, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Andrea
authors.initials: A
authors.lastname: Sacchetti
authors.affiliation: Dept. of Pathology, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Johan M
authors.initials: JM
authors.lastname: Kros
authors.affiliation: Dept. of Neurology, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Peter
authors.initials: P
authors.lastname: Sillevis Smitt
authors.affiliation: Dept. of Pulmonary diseases, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Joachim
authors.initials: J
authors.lastname: Aerts
authors.affiliation: Dept. of Neurology, Erasmus Medical Center, Rotterdam, The Netherlands. Electronic address: p.french@erasmusmc.nl.
authors.forename: Pim J
authors.initials: PJ
authors.lastname: French
mesh_terms: D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000235:genetics; D066246:ErbB Receptors / Q000235:genetics / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D009154:Mutation; D040081:Protein Array Analysis / Q000379:methods; D011485:Protein Binding / Q000235:genetics; D015398:Signal Transduction / Q000235:genetics; D014162:Transfection
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C512478:EGFR protein, human; D066246:ErbB Receptors
keywords: Cancer; DOCK4; EGFR; Glioma; Pulmonary adenocarcinoma