Farrell, JenniferJenniferFarrellKelly, CiaraCiaraKellyRauch, JensJensRauchKida, KatarzynaKatarzynaKidaGarcia Munoz, AmayaAmayaGarcia MunozMonsefi, NaserNaserMonsefiTurriziani, BenedettaBenedettaTurrizianiDoherty, CarolanneCarolanneDohertyMehta, J. P.J. P.MehtaMatallanas, DavidDavidMatallanasSimpson, Jeremy C.Jeremy C.SimpsonKolch, WalterWalterKolchKriegsheim, Alexander vonAlexander vonKriegsheim2016-04-042016-04-042014 Ameri2014-06-06Journal of Proteome Researchhttp://hdl.handle.net/10197/7540Epithelial to mesenchymal transition (EMT) is a fundamental cell differentiation/dedifferentiation process which is associated with dramatic morphological changes. Formerly polarized and immobile epithelial cells which form cell junctions and cobblestone-like cell sheets undergo a transition into highly motile, elongated, mesenchymal cells lacking cell-to-cell adhesions. To explore how the proteome is affected during EMT we profiled protein expression and tracked cell biological markers in Madin-Darby kidney epithelial cells undergoing hepatocyte growth factor (HGF) induced EMT. We were able to identify and quantify over 4000 proteins by mass spectrometry. Enrichment analysis of this revealed that expression of proteins associated with the ubiquitination machinery was induced, whereas expression of proteins regulating apoptotic pathways was suppressed. We show that both the mammalian Hippo/MST2 and the ISG15 pathways are regulated at the protein level by ubiquitin ligases. Inhibition of the Hippo pathway by overexpression of either ITCH or A-Raf promotes HGF-induced EMT. Conversely, ISG15 overexpression is sufficient to induce cell scattering and an elongated morphology without external stimuli. Thus, we demonstrate for the first time that the Hippo/MST2 and ISG15 pathways are regulated during growth-factor induced EMT.enThis document is the unedited author's version of a Submitted Work that was subsequently accepted for publication in Journal of Proteome Research, copyright © American Chemical Society after peer review. To access the final edited and published work, see http://pubs.acs.org/doi/abs/10.1021/pr5000285.Epithelial mesenchymal transitionHippo-pathwayISG5SILACQuantitative expression proteomicsYAP1Journal Article1362874288610.1021/pr50002852016-02-22https://creativecommons.org/licenses/by-nc-nd/3.0/ie/