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Nuclear extracellular signal-regulated kinase 2 phosphorylates p53 at Thr55 in response to doxorubicin

P Y Yeh1, S E Chuang, K H Yeh

  • 1Cancer Research Center, College of Medicine, Taipei, Taiwan, Republic of China.

Biochemical and Biophysical Research Communications
|June 21, 2001
PubMed

Insights

Nuclear ERK2 phosphorylates p53 at Thr55, impacting its transactivating activity. This interaction suggests ERK2

Area of Science:

  • Molecular Biology
  • Cellular Signaling
  • Cancer Research

Background:

  • The tumor suppressor protein p53 plays a critical role in cellular responses to DNA damage.
  • Extracellular signal-regulated kinases (ERK) are key components of signaling pathways involved in cell growth and survival.
  • The precise mechanisms by which ERK signaling influences p53 function, particularly in response to genotoxic stress, remain incompletely understood.

Purpose of the Study:

  • To investigate the interaction between ERK2 and p53.
  • To identify the specific site of p53 phosphorylation by ERK2.
  • To determine the functional consequence of this phosphorylation on p53 activity.

Main Methods:

  • Co-immunoprecipitation and Western blotting to confirm physical interaction between ERK2 and p53.
  • Site-directed mutagenesis of potential phosphorylation sites in the N-terminal domain of p53.
  • In vitro kinase assays using recombinant proteins to identify the phosphorylation site.
  • Electromobility shift assays to assess the impact of phosphorylation on p53 transactivating activity.

Main Results:

  • Nuclear ERK2 directly phosphorylates p53 at Threonine 55 (Thr55).
  • ERK2 physically interacts with p53, confirmed by co-immunoprecipitation.
  • Phosphorylation of p53 by ERK2 at Thr55 modulates its transactivating function.

Conclusions:

  • ERK2 directly phosphorylates p53 at Thr55 in response to DNA damage.
  • This phosphorylation event influences p53's ability to transactivate target genes.
  • ERK2-mediated p53 phosphorylation represents a novel mechanism in the cellular response to DNA damage.

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