Oncogenic Mutations Rewire Signaling Pathways by Switching Protein Recruitment to Phosphotyrosine Sites

TY - JOUR

T1 - Oncogenic Mutations Rewire Signaling Pathways by Switching Protein Recruitment to Phosphotyrosine Sites

AU - Lundby, Alicia

AU - Franciosa, Giulia

AU - Emdal, Kristina B

AU - Refsgaard, Jan C

AU - Gnosa, Sebastian P

AU - Bekker-Jensen, Dorte B

AU - Secher, Anna

AU - Maurya, Svetlana R

AU - Paul, Indranil

AU - Mendez, Blanca L

AU - Kelstrup, Christian D

AU - Francavilla, Chiara

AU - Kveiborg, Marie

AU - Montoya, Guillermo

AU - Jensen, Lars J

AU - Olsen, Jesper V

N1 - Copyright © 2019 Elsevier Inc. All rights reserved.

PY - 2019/10/3

Y1 - 2019/10/3

N2 - Tyrosine phosphorylation regulates multi-layered signaling networks with broad implications in (patho)physiology, but high-throughput methods for functional annotation of phosphotyrosine sites are lacking. To decipher phosphotyrosine signaling directly in tissue samples, we developed a mass-spectrometry-based interaction proteomics approach. We measured the in vivo EGF-dependent signaling network in lung tissue quantifying >1,000 phosphotyrosine sites. To assign function to all EGF-regulated sites, we determined their recruited protein signaling complexes in lung tissue by interaction proteomics. We demonstrated how mutations near tyrosine residues introduce molecular switches that rewire cancer signaling networks, and we revealed oncogenic properties of such a lung cancer EGFR mutant. To demonstrate the scalability of the approach, we performed >1,000 phosphopeptide pulldowns and analyzed them by rapid mass spectrometric analysis, revealing tissue-specific differences in interactors. Our approach is a general strategy for functional annotation of phosphorylation sites in tissues, enabling in-depth mechanistic insights into oncogenic rewiring of signaling networks.

AB - Tyrosine phosphorylation regulates multi-layered signaling networks with broad implications in (patho)physiology, but high-throughput methods for functional annotation of phosphotyrosine sites are lacking. To decipher phosphotyrosine signaling directly in tissue samples, we developed a mass-spectrometry-based interaction proteomics approach. We measured the in vivo EGF-dependent signaling network in lung tissue quantifying >1,000 phosphotyrosine sites. To assign function to all EGF-regulated sites, we determined their recruited protein signaling complexes in lung tissue by interaction proteomics. We demonstrated how mutations near tyrosine residues introduce molecular switches that rewire cancer signaling networks, and we revealed oncogenic properties of such a lung cancer EGFR mutant. To demonstrate the scalability of the approach, we performed >1,000 phosphopeptide pulldowns and analyzed them by rapid mass spectrometric analysis, revealing tissue-specific differences in interactors. Our approach is a general strategy for functional annotation of phosphorylation sites in tissues, enabling in-depth mechanistic insights into oncogenic rewiring of signaling networks.

U2 - 10.1016/j.cell.2019.09.008

DO - 10.1016/j.cell.2019.09.008

M3 - Journal article

C2 - 31585087

SN - 0092-8674

VL - 179

SP - 543

EP - 560

JO - Cell

JF - Cell

IS - 2

ER -