Deciphering MET-dependent modulation of global cellular responses to DNA damage by quantitative phosphoproteomics.

Abstract

Increasing evidence suggests that interference with growth factor receptor tyrosine kinase (RTK) signaling can affect DNA damage response (DDR) networks, with a consequent impact on cellular responses to DNA-damaging agents widely used in cancer treatment. In that respect, the MET RTK is deregulated in abundance and/or activity in a variety of human tumors. Using two proteomic techniques, we explored how disrupting MET signaling modulates global cellular phosphorylation response to ionizing radiation (IR). Following an immunoaffinity-based phosphoproteomic discovery survey, we selected candidate phosphorylation sites for extensive characterization by targeted proteomics focusing on phosphorylation sites in both signaling networks. Several substrates of the DDR were confirmed to be modulated by sequential MET inhibition and IR, or MET inhibition alone. Upon combined treatment, for two substrates, NUMA1 S395 and CHEK1 S345, the gain and loss of phosphorylation, respectively, were recapitulated using invivo tumor models by immunohistochemistry, with possible utility in future translational research. Overall, we have corroborated phosphorylation sites at the intersection between MET and the DDR signaling networks, and suggest that these represent a class of proteins at the interface between oncogene-driven proliferation and genomic stability.

• Authors: Ariel Bensimon, Jonas P Koch, Paola Francica, Selina M Roth, Rahel Riedo, Astrid A Glück, Eleonora Orlando, Andree Blaukat, Daniel M Aebersold, Yitzhak Zimmer, Ruedi Aebersold, Michaela Medová
• Journal: Molecular oncology (Mol Oncol) Vol. 14 Issue 6 Pg. 1185-1206 (06 2020) ISSN: 1878-0261 [Electronic] United States
• PMID: 32336009 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2020 The Authors. Published by FEBS Press and John Wiley & Sons Ltd.

Chemical References

• Neoplasm Proteins
• Phosphoproteins

Topics

• Animals
• Cell Line, Tumor
• DNA Damage
• DNA Repair (radiation effects)
• Down-Regulation (radiation effects)
• Epithelium (pathology, radiation effects)
• Female
• Humans
• Mesoderm (pathology, radiation effects)
• Mice
• Neoplasm Proteins (metabolism)
• Phosphatidylinositol 3-Kinases (metabolism)
• Phosphoproteins (metabolism)
• Phosphorylation (radiation effects)
• Proteomics
• Radiation, Ionizing
• Reproducibility of Results
• Substrate Specificity (radiation effects)
• Xenograft Model Antitumor Assays