摘要：SummaryMyogenesis is governed by signaling networks that are tightly regulated in a time-dependent manner. Although different protein kinases have been identified, knowledge of the global signaling networks and their downstream substrates during myogenesis remains incomplete. Here, we map the myogenic differentiation of C2C12 cells using phosphoproteomics and proteomics. From these data, we infer global kinase activity and predict the substrates that are involved in myogenesis. We found that multiple mitogen-activated protein kinases (MAPKs) mark the initial wave of signaling cascades. Further phosphoproteomic and proteomic profiling with MAPK1/3 and MAPK8/9 specific inhibitions unveil their shared and distinctive roles in myogenesis. Lastly, we identified and validated the transcription factor nuclear factor 1 X-type (NFIX) as a novel MAPK1/3 substrate and demonstrated the functional impact of NFIX phosphorylation on myogenesis. Altogether, these data characterize the dynamics, interactions, and downstream control of kinase signaling networks during myogenesis on a global scale.Graphical abstractDisplay OmittedHighlights•Phosphoproteomic and proteomic maps of myogenic differentiation of C2C12 cells•Myogenic kinome activity and kinase-substrates prediction using machine learning•MAPK1/3 and MAPK8/9 inhibition unveil shared and distinctive effects on myogenesis•Validation of NFIX phosphorylation by MAPK1/3 and its impact on myogenesisCell biology; Developmental biology; Proteomics
