A Protein Turnover Signaling Motif Controls the Stimulus-Sensitivity of Stress Response Pathways

Stimulus-induced perturbations from the steady state are a hallmark of signal transduction. In some signaling modules, the steady state is characterized by rapid synthesis and degradation of signaling proteins. Conspicuous among these are the p53 tumor suppressor, its negative regulator Mdm2, and the negative feedback regulator of NFκB, IκBα. We investigated the physiological importance of this turnover, or flux, using a computational method that allows flux to be systematically altered independently of the steady state protein abundances. Applying our method to a prototypical signaling module, we show that flux can precisely control the dynamic response to perturbation. Next, we applied our method to experimentally validated models of p53 and NFκB signaling. We find that high p53 flux is required for oscillations in response to a saturating dose of ionizing radiation (IR). In contrast, high flux of Mdm2 is not required for oscillations but preserves p53 sensitivity to sub-saturating doses of IR. In the NFκB system, degradation of NFκB-bound IκB by the IκB kinase (IKK) is required for activation in response to TNF, while high IKK-independent degradation prevents spurious activation in response to metabolic stress or low doses of TNF. Our work identifies flux pairs with opposing functional effects as a signaling motif that controls the stimulus-sensitivity of the p53 and NFκB stress-response pathways, and may constitute a general design principle in signaling pathways. Eukaryotic cells constantly synthesize new proteins and degrade old ones. While most proteins are degraded within 24 hours of being synthesized, some proteins are short-lived and exist for only minutes. Using mathematical models, we asked how rapid turnover, or flux, of signaling proteins might regulate the activation of two well-known transcription factors, p53 and NFκB. p53 is a cell cycle regulator that is activated in response to DNA damage, for example, due to ionizing radiation. NFκB is a regulator of immunity and responds to inflammatory signals like the macrophage-secreted cytokine, TNF. Both p53 and NFκB are controlled by at least one flux whose effect on activation is positive and one whose effect is negative. For p53 these are the turnover of p53 and Mdm2, respectively. For NFκB they are the TNF-dependent and -independent turnover of the NFκB inhibitor, IκB. We find that juxtaposition of a positive and negative flux allows for precise tuning of the sensitivity of these transcription factors to different environmental signals. Our results therefore suggest that rapid synthesis and degradation of signaling proteins, though energetically wasteful, may be a common mechanism by which eukaryotic cells regulate their sensitivity to environmental stimuli.