title: USP9X Controls EGFR Fate by Deubiquitinating the Endocytic Adaptor Eps15.
abstract: Following activation by its cognate ligand(s), the epidermal growth factor receptor (EGFR) is rapidly routed to the lysosome for degradation in a ubiquitination-dependent fashion. This pathway represents the major mechanism of long-term attenuation of EGFR signaling, and its deregulation is a significant feature in different types of cancers. Here we demonstrate, through a systematic RNAi-based approach, that several deubiquitinating (DUB) enzymes extend or decrease EGFR half-life upon EGF stimulation. We focus on USP9X, whose depletion severely affects EGFR turnover, interfering with its internalization and trafficking. We identify the endocytic protein Eps15 as one of the critical substrates of USP9X, and we map the Eps15 ubiquitination sites. We found that Eps15 monoubiquitination occurs already at minimal dose of EGF stimulation and is essential for EGFR internalization. Overall, our findings identify USP9X as a novel regulator of EGFR endocytosis and suggest a model whereby cycles of ubiquitination and deubiquitination events on endocytic accessory proteins may regulate the internalization and trafficking of the EGFR toward the lysosomes.
authors.affiliation: IFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy; DIPO, Dipartimento di Oncologia ed Emato-Oncologia, Università degli Studi di Milano, Via di Rudinì 8, 20122 Milan, Italy.
authors.forename: Michol Giovanna
authors.initials: MG
authors.lastname: Savio
authors.affiliation: IFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy.
authors.forename: Nadine
authors.initials: N
authors.lastname: Wollscheid
authors.affiliation: IFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Cavallaro
authors.affiliation: IFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy.
authors.forename: Veronica
authors.initials: V
authors.lastname: Algisi
authors.affiliation: IFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy; DIPO, Dipartimento di Oncologia ed Emato-Oncologia, Università degli Studi di Milano, Via di Rudinì 8, 20122 Milan, Italy; IEO, Istituto Europeo di Oncologia, Via Ripamonti 435, 20141 Milan, Italy.
authors.forename: Pier Paolo
authors.initials: PP
authors.lastname: Di Fiore
authors.affiliation: IFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Sigismund
authors.affiliation: IFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Maspero
authors.affiliation: IFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy; DIPO, Dipartimento di Oncologia ed Emato-Oncologia, Università degli Studi di Milano, Via di Rudinì 8, 20122 Milan, Italy. Electronic address: simona.polo@ifom.eu.
authors.forename: Simona
authors.initials: S
authors.lastname: Polo
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D045744:Cell Line, Tumor; D004705:Endocytosis / Q000502:physiology; D011992:Endosomes / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism*; D006801:Humans; D008247:Lysosomes / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000502:physiology; D021381:Protein Transport / Q000502:physiology; D043222:Ubiquitin Thiolesterase / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C486206:EPS15 protein, human; C506234:USP9X protein, human; D004815:Epidermal Growth Factor; C512478:EGFR protein, human; D066246:ErbB Receptors; D043222:Ubiquitin Thiolesterase
keywords: EGFR; Eps15; USP9X; deubiquitinating enzymes; ubiquitin