A systematic approach to decipher crosstalk in the p53 signaling pathway using single cell dynamics

PLoS Comput Biol. 2020 Jun 26;16(6):e1007901. doi: 10.1371/journal.pcbi.1007901. eCollection 2020 Jun.


The transcription factors NF-κB and p53 are key regulators in the genotoxic stress response and are critical for tumor development. Although there is ample evidence for interactions between both networks, a comprehensive understanding of the crosstalk is lacking. Here, we developed a systematic approach to identify potential interactions between the pathways. We perturbed NF-κB signaling by inhibiting IKK2, a critical regulator of NF-κB activity, and monitored the altered response of p53 to genotoxic stress using single cell time lapse microscopy. Fitting subpopulation-specific computational p53 models to this time-resolved single cell data allowed to reproduce in a quantitative manner signaling dynamics and cellular heterogeneity for the unperturbed and perturbed conditions. The approach enabled us to untangle the integrated effects of IKK/ NF-κB perturbation on p53 dynamics and thereby derive potential interactions between both networks. Intriguingly, we find that a simultaneous perturbation of multiple processes is necessary to explain the observed changes in the p53 response. Specifically, we show interference with the activation and degradation of p53 as well as the degradation of Mdm2. Our results highlight the importance of the crosstalk and its potential implications in p53-dependent cellular functions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Microscopy / methods
  • NF-kappa B / metabolism
  • Reproducibility of Results
  • Signal Transduction
  • Single-Cell Analysis*
  • Tumor Suppressor Protein p53 / physiology*


  • NF-kappa B
  • Tumor Suppressor Protein p53

Grant support

This work was supported by German Cancer Aid (project number 111645 to A.L.). FK was funded by a PhD fellowship of the graduate school ‘Computational Systems Biology (CSB)’ of the German Research Foundation (DFG-Graduiertenkolleg 1772). The project was supported by a grant from the German Federal Ministry of Education and Research BMBF (Project ProSiTu, 0316047A) and the Personalized Medicine Initiative ‘iMed’ of the Helmholtz Association awarded to JW. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.