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DUSP6/MKP-3 inactivates ERK1/2 but fails to bind and inactivate ERK5

Rebecca S Arkell1, Robin J Dickinson, Matthew Squires

  • 1Laboratory of Molecular Signalling, The Babraham Institute, Babraham Research Campus, Cambridge, CB22 3AT, UK.

Cellular Signalling
|February 19, 2008
PubMed

Insights

Dual-specificity phosphatases (DUSP) inactivate protein kinases. This study confirms DUSP6 specifically targets extracellular signal-regulated kinase-1 and -2 (ERK1/2), not ERK5, highlighting its role in regulating ERK1/2 signaling pathways.

Area of Science:

  • Molecular Biology
  • Cell Signaling
  • Enzymology

Background:

  • Extracellular signal-regulated kinase-1 and -2 (ERK1/2) and ERK5 are protein kinases activated by phosphorylation.
  • Dual-specificity protein phosphatases (DUSP) inactivate these kinases by de-phosphorylating a conserved TEY motif.
  • DUSP6 is a cytoplasmic DUSP implicated in ERK1/2 regulation, with some evidence suggesting potential cross-reactivity with ERK5.

Purpose of the Study:

  • To investigate and clarify the substrate specificity of DUSP6 towards ERK1/2 and ERK5.
  • To determine if DUSP6 can inactivate ERK5, despite its known role in regulating ERK1/2.

Main Methods:

  • Yeast and human cell-based binding assays to assess DUSP6-kinase interactions.
  • In vitro kinase-phosphatase assays using recombinant ERK2 and ERK5 with DUSP6.
  • Functional assays involving ectopic DUSP6 expression to examine de-phosphorylation and downstream signaling effects (MEK1/ERK1/2 and MEK5/ERK5 pathways).

Main Results:

  • DUSP6 demonstrated binding to ERK1/2 but not to ERK5 in both yeast and human cells.
  • Recombinant ERK2, but not ERK5, catalytically activated DUSP6.
  • Ectopic DUSP6 expression de-phosphorylated ERK2 but not ERK5, and specifically inhibited ERK1/2-mediated transcription (GAL4-ELK1) without affecting ERK5-mediated transcription (GAL4-MEF2D).

Conclusions:

  • DUSP6 exclusively targets and inactivates ERK1/2.
  • Even at high expression levels, DUSP6 does not affect ERK5 activity, confirming its high specificity for the ERK1/2 pathway.
  • These findings solidify DUSP6's role as an ERK1/2-specific regulator.

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