Multilayered proteomics reveals molecular switches dictating ligand-dependent EGFR trafficking

Chiara Francavilla, Moreno Papetti, Kristoffer T G Rigbolt, Anna-Kathrine Pedersen, Jon O Sigurdsson, Giuseppe Cazzamali, Gopal Karemore, Blagoy Blagoev, Jesper V Olsen

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

A fascinating conundrum in cell signaling is how stimulation of the same receptor tyrosine kinase with distinct ligands generates specific outcomes. To decipher the functional selectivity of EGF and TGF-α, which induce epidermal growth factor receptor (EGFR) degradation and recycling, respectively, we devised an integrated multilayered proteomics approach (IMPA). We analyzed dynamic changes in the receptor interactome, ubiquitinome, phosphoproteome, and late proteome in response to both ligands in human cells by quantitative MS and identified 67 proteins regulated at multiple levels. We identified RAB7 phosphorylation and RCP recruitment to EGFR as switches for EGF and TGF-α outputs, controlling receptor trafficking, signaling duration, proliferation, and migration. By manipulating RCP levels or phosphorylation of RAB7 in EGFR-positive cancer cells, we were able to switch a TGF-α-mediated response to an EGF-like response or vice versa as EGFR trafficking was rerouted. We propose IMPA as an approach to uncover fine-tuned regulatory mechanisms in cell signaling.

Original languageEnglish
JournalNature Structural and Molecular Biology
Volume23
Issue number6
Pages (from-to)608-18
Number of pages11
ISSN1545-9993
DOIs
Publication statusPublished - Jun 2016

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