YES oncogenic activity is specified by its SH4 domain and regulates RAS/MAPK signaling in colon carcinoma cells.
Members of the SRC family of tyrosine kinases (SFK) display important functions in human cancer, but their specific role in tumorigenesis remains unclear. We previously demonstrated that YES regulates a unique oncogenic signaling important for colorectal cancer (CRC) progression that is not shared with SRC. Here, we addressed the underlying mechanism involved in this process. We show that YES oncogenic signaling relies on palmitoylation of its SH4 domain that controls YES localization in cholesterol-enriched membrane micro-domains. Specifically, deletion of the palmitoylation site compromised YES transforming activity, while addition of a palmitoylation site in the SH4 domain of SRC was sufficient for SRC to restore the transforming properties of cells in which YES had been silenced. Subsequently, SILAC phosphoproteomic analysis revealed that micro-domain-associated cell adhesive components and receptor tyrosine kinases are major YES substrates. YES also phosphorylates upstream regulators of RAS/MAPK signaling, including EGFR, SHC and SHP2, which were not targeted by SRC due to the absence of palmitoylation. Accordingly, EGFR-induced MAPK activity was attenuated by YES down-regulation, while increased RAS activity significantly restored cell transformation that was lost upon YES silencing. Collectively, these results uncover a critical role for the SH4 domain in the specification of SFK oncogenic activity and a selective role for YES in the induction of RAS/MAPK signaling in CRC cells.
CNRS UMR5237, University of Montpellier, CRBM 1919 Route de Mende, 34000 Montpellier, France ; CNRS, UMR5203 CNRS, INSERM U661 University of Montpellier, IGF 34000 Montpellier, France.
Fanny
F
Dubois
CNRS UMR5237, University of Montpellier, CRBM 1919 Route de Mende, 34000 Montpellier, France ; Novartis CH-4002 Basel, Switzerland.
Cédric
C
Leroy
CNRS UMR5237, University of Montpellier, CRBM 1919 Route de Mende, 34000 Montpellier, France.
Valérie
V
Simon
CNRS UMR5237, University of Montpellier, CRBM 1919 Route de Mende, 34000 Montpellier, France ; CNRS, UMR5048 CNRS, INSERM U1048, University of Montpellier, CBS 34000 Montpellier, France.
Christine
C
Benistant
CNRS UMR5237, University of Montpellier, CRBM 1919 Route de Mende, 34000 Montpellier, France.
Serge
S
Roche
SRC; YES; colon cancer; oncogenic signaling; phosphoproteomics; tyrosine kinase