Global view of the RAF-MEK-ERK module and its immediate downstream effectors.

Autor: Santini CC; Technical University of Denmark (DTU), Kgs. Lyngby, 2800, Denmark.; Celgene Institute Translational Research Europe (CITRE), Seville, E-41092, Spain., Longden J; Biotech Research & Innovation Centre, University of Copenhagen, Copenhagen, 2200, Denmark., Schoof EM; Technical University of Denmark (DTU), Kgs. Lyngby, 2800, Denmark., Simpson CD; Biotech Research & Innovation Centre, University of Copenhagen, Copenhagen, 2200, Denmark., Jeschke GR; Department of Pharmacology, Yale University, New Haven, 06520, USA., Creixell P; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, USA., Kim J; Samsung Genome Institute, Samsung medical Center, Seoul, 06351, South Korea., Wu X; Icahn School of Medicine at Mount Sinai, New York, 10029-5674, USA., Turk BE; Department of Pharmacology, Yale University, New Haven, 06520, USA., Rosen N; Memorial Sloan Kettering Cancer Center, New York, 10065, USA., Poulikakos PI; Icahn School of Medicine at Mount Sinai, New York, 10029-5674, USA., Linding R; Technical University of Denmark (DTU), Kgs. Lyngby, 2800, Denmark. linding@lindinglab.org.; Biotech Research & Innovation Centre, University of Copenhagen, Copenhagen, 2200, Denmark. linding@lindinglab.org.; Institute of Biology, Humboldt-Universität zu Berlin, Berlin, 10115, Germany. linding@lindinglab.org.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2019 Jul 26; Vol. 9 (1), pp. 10865. Date of Electronic Publication: 2019 Jul 26.
DOI: 10.1038/s41598-019-47245-x
Abstrakt: Small molecule inhibitors of BRAF and MEK have proven effective at inhibiting tumor growth in melanoma patients, however this efficacy is limited due to the almost universal development of drug resistance. To provide advanced insight into the signaling responses that occur following kinase inhibition we have performed quantitative (phospho)-proteomics of human melanoma cells treated with either dabrafenib, a BRAF inhibitor; trametinib, a MEK inhibitor or SCH772984, an ERK inhibitor. Over nine experiments we identified 7827 class I phosphorylation sites on 4960 proteins. This included 54 phosphorylation sites that were significantly down-modulated after exposure to all three inhibitors, 34 of which have not been previously reported. Functional analysis of these novel ERK targets identified roles for them in GTPase activity and regulation, apoptosis and cell-cell adhesion. Comparison of the results presented here with previously reported phosphorylation sites downstream of ERK showed a limited degree of overlap suggesting that ERK signaling responses may be highly cell line and cue specific. In addition we identified 26 phosphorylation sites that were only responsive to dabrafenib. We provide further orthogonal experimental evidence for 3 of these sites in human embryonic kidney cells over-expressing BRAF as well as further computational insights using KinomeXplorer. The validated phosphorylation sites were found to be involved in actin regulation, which has been proposed as a novel mechanism for inhibiting resistance development. These results would suggest that the linearity of the BRAF-MEK-ERK module is at least context dependent.
Databáze: MEDLINE
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