Endogenous, regulatory cysteine sulfenylation of ERK kinases in response to proliferative signals

Free Radic Biol Med. 2017 Nov:112:534-543. doi: 10.1016/j.freeradbiomed.2017.08.018. Epub 2017 Aug 24.

Abstract

ERK-dependent signaling is key to many pathways through which extracellular signals are transduced into cell-fate decisions. One conundrum is the way in which disparate signals induce specific responses through a common, ERK-dependent kinase cascade. While studies have revealed intricate ways of controlling ERK signaling through spatiotemporal localization and phosphorylation dynamics, additional modes of ERK regulation undoubtedly remain to be discovered. We hypothesized that fine-tuning of ERK signaling could occur by cysteine oxidation. We report that ERK is actively and directly oxidized by signal-generated H2O2 during proliferative signaling, and that ERK oxidation occurs downstream of a variety of receptor classes tested in four cell lines. Furthermore, within the tested cell lines and proliferative signals, we observed that both activation loop-phosphorylated and non-phosphorylated ERK undergo sulfenylation in cells and that dynamics of ERK sulfenylation is dependent on the cell growth conditions prior to stimulation. We also tested the effect of endogenous ERK oxidation on kinase activity and report that phosphotransfer reactions are reversibly inhibited by oxidation by as much as 80-90%, underscoring the importance of considering this additional modification when assessing ERK activation in response to extracellular signals.

Keywords: Growth factor signaling; Kinase; Reactive cysteine, ERK, MAPK; Redox regulation; Sulfenic acid.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cell Line, Tumor
  • Cysteine / metabolism
  • Epithelial Cells / cytology
  • Epithelial Cells / drug effects*
  • Epithelial Cells / metabolism
  • Extracellular Signal-Regulated MAP Kinases / genetics
  • Extracellular Signal-Regulated MAP Kinases / metabolism*
  • Fibroblasts / cytology
  • Fibroblasts / drug effects*
  • Fibroblasts / metabolism
  • Humans
  • Hydrogen Peroxide / pharmacology
  • MAP Kinase Signaling System
  • Mice
  • NIH 3T3 Cells
  • Oxidation-Reduction
  • Protein Processing, Post-Translational*
  • Sulfenic Acids / metabolism*

Substances

  • Sulfenic Acids
  • Hydrogen Peroxide
  • Extracellular Signal-Regulated MAP Kinases
  • Cysteine