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ERK oscillation-dependent gene expression patterns and deregulation by stress response.

Katrina M Waters1, Brian S Cummings, Harish Shankaran

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Extracellular signal-regulated kinase (ERK) oscillations control specific genes in human skin cells. The p38 stress response disrupts these oscillations, impacting gene regulation and cellular responses to carcinogens like bromate.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Signal Transduction

Background:

  • Extracellular signal-regulated kinase (ERK) pathway oscillations are increasingly recognized for their role in cellular processes.
  • The interplay between ERK oscillations and stress response pathways, such as p38, is not fully understood.
  • Understanding these interactions is crucial for deciphering cellular responses to environmental stimuli and carcinogens.

Purpose of the Study:

  • To investigate if ERK oscillations regulate a unique gene subset in human keratinocytes.
  • To determine if the p38 stress response pathway inhibits ERK oscillations.
  • To elucidate the role of Mediator Complex Subunit 1 (MED1) in ERK oscillation-dependent gene expression.

Main Methods:

  • DNA microarray analysis to identify oscillation-unique genes.
  • Network reconstruction to predict key regulatory nodes.
  • Western blotting to assess protein phosphorylation (e.g., phospho-MED1, phospho-p38).
  • Pharmacological inhibition of p38 using SB203580 and SB202190.

Main Results:

  • DNA microarray identified numerous genes uniquely regulated by ERK oscillations.
  • MED1 was predicted and validated as a key mediator of oscillation-dependent gene expression, with increased phosphorylation in oscillating cells.
  • p38 inhibition enhanced ERK oscillation amplitude and MED1 phosphorylation.
  • Bromate, a carcinogen activating p38, inhibited ERK oscillations and decreased MED1 phosphorylation in various cell types.
  • Bromate-induced decrease in MED1 phosphorylation was reversible with p38 inhibitors.

Conclusions:

  • ERK pathway oscillations significantly alter homeostatic gene regulation patterns in human keratinocytes.
  • The p38 stress response pathway directly interferes with ERK oscillations and MED1 regulation.
  • Cellular responses to perturbations, such as carcinogen exposure, differ between oscillating and non-oscillating cells, highlighting the importance of dynamic signaling.
  • These findings provide insights into how environmental factors can disrupt normal cellular signaling and gene expression.