Coordination of MAPK and p53 dynamics in the cellular responses to DNA damage and oxidative stress

Abstract

In response to different cellular stresses, the transcription factor p53 undergoes different dynamics. p53 dynamics, in turn, control cell fate. However, distinct stresses can generate the same p53 dynamics but different cell fate outcomes, suggesting integration of dynamic information from other pathways is important for cell fate regulation. To determine how MAPK activities affect p53‐mediated responses to DNA breaks and oxidative stress, we simultaneously tracked p53 and either ERK, JNK, or p38 activities in single cells. While p53 dynamics were comparable between the stresses, cell fate outcomes were distinct. Combining MAPK dynamics with p53 dynamics was important for distinguishing between the stresses and for generating temporal ordering of cell fate pathways. Furthermore, cross‐talk between MAPKs and p53 controlled the balance between proliferation and cell death. These findings provide insight into how cells integrate signaling pathways with distinct temporal patterns of activity to encode stress specificity and drive different cell fate decisions.

Temporal Coordination of the Transcription Factor Response to H2O2stress

“…We then treated cells containing both reporters with four different concentrations of H2O2 and measured TF levels in single cells every 20 minutes for 24 hours. At the lowest H2O2 dose (50 µM), FOXO1 remained largely inactive (in the cytoplasm), while p53 levels oscillated, as shown previously in response to DNA double strand breaks and H2O2 40,41 (Figure 2A, 2B, Supp Movie 1). At higher H2O2 concentrations, FOXO1 accumulated in the nucleus within 1 hour of treatment in a subset of cells (Figure 2A,C-E, Supp Movies 2,3,4).…”

“…While other cells show large bursts of p53 levels similar to the response to UV irradiation 42 . The proportion of cells with oscillating p53 levels decreased with dose as shown by autocorrelation analysis, and recently observed in retinal pigment epithelial cells 40 (Figure S2A-D). These differences correlated with cell survival as the p53 levels in cells that died reached higher levels than those that survived (Figure S2G and S2H).…”

“…This cell line also contains a H2B-CFP tag for tracking nuclei 39 . For p53, we added an exogenously expressed p53-mCherry reporter used and validated in previous studies 40 . We then treated cells containing both reporters with four different concentrations of H 2 O 2 and measured TF levels in single cells every 20 minutes for 24 hours.…”

Temporal Coordination of the Transcription Factor Response to H2O2 stress

“…This cell line also contains a H2B-CFP tag for tracking nuclei 36 . For p53, we added an exogenously expressed p53-mCherry reporter used and validated in previous studies 37 . We then treated cells containing both reporters with four different concentrations of H 2 O 2 and measured TF levels in single cells every 20 minutes for 24 hours.…”

YAP localization mediates mechanical adaptation of human cancer cells during extravasationin vivo

“…To establish MDA-MB-231 and A375SM cell lines expressing H2B-mCerulean3, cells were transduced with lentivirus expressing pLentiPGK Hygro DEST H2B-mCerulean3 (Addgene: 90234) diluted 1:50 in growth medium supplemented with 8 μg/ml protamine sulfate and 10 μM HEPES. After 72 hours, media was removed and selection was performed in growth medium supplemented with 500 μg/ml hygromycin [72]. Positive cells were isolated via Fluorescence-Activated Cell Sorting (FACS).…”