PUBLICATION
Oncogenic Mutations Rewire Signaling Pathways by Switching Protein Recruitment to Phosphotyrosine Sites
- Authors
- Lundby, A., Franciosa, G., Emdal, K.B., Refsgaard, J.C., Gnosa, S.P., Bekker-Jensen, D.B., Secher, A., Maurya, S.R., Paul, I., Mendez, B.L., Kelstrup, C.D., Francavilla, C., Kveiborg, M., Montoya, G., Jensen, L.J., Olsen, J.V.
- ID
- ZDB-PUB-191005-12
- Date
- 2019
- Source
- Cell 179: 543-560.e26 (Journal)
- Registered Authors
- Keywords
- EGFR, SH2, cancer signaling, interaction proteomics, mass spectrometry, peptide pulldown, phosphopeptides, phosphoproteomics, phosphotyrosine, zebrafish
- MeSH Terms
-
- A549 Cells
- Animals
- Carcinogenesis/genetics*
- ErbB Receptors/genetics*
- Humans
- Lung Neoplasms/genetics*
- Lung Neoplasms/metabolism*
- Mass Spectrometry/methods
- Mutation
- Phosphoproteins/metabolism
- Phosphorylation
- Phosphotyrosine/metabolism*
- Proteomics
- Rats
- Rats, Sprague-Dawley
- Zebrafish
- PubMed
- 31585087 Full text @ Cell
Citation
Lundby, A., Franciosa, G., Emdal, K.B., Refsgaard, J.C., Gnosa, S.P., Bekker-Jensen, D.B., Secher, A., Maurya, S.R., Paul, I., Mendez, B.L., Kelstrup, C.D., Francavilla, C., Kveiborg, M., Montoya, G., Jensen, L.J., Olsen, J.V. (2019) Oncogenic Mutations Rewire Signaling Pathways by Switching Protein Recruitment to Phosphotyrosine Sites. Cell. 179:543-560.e26.
Abstract
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping