Prostaglandin A2-mediated stabilization of p21 mRNA through an ERK-dependent pathway requiring the RNA-binding protein HuR

J Biol Chem. 2004 Nov 19;279(47):49298-306. doi: 10.1074/jbc.M407535200. Epub 2004 Sep 14.

Abstract

Treatment with the stress agent prostaglandin A2 (PGA2) induces expression of the cyclin-dependent kinase inhibitor p21. Here, we present evidence that p21 expression increases through PGA2-triggered stabilization of the p21 mRNA and further show that these events require the mitogen-activated protein (MAP) kinase ERK. Binding experiments using either endogenous p21 mRNA or in vitro-labeled p21 transcripts revealed a specific PGA2-dependent association of the p21 mRNA with the RNA-binding protein HuR. Interestingly, although inhibition of the ERK pathway did not prevent the PGA2-triggered increase in cytoplasmic HuR, it did impair the formation of endogenous and in vitro [HuR-p21 mRNA] complexes and further prevented the PGA2-mediated stabilization of the p21 mRNA, suggesting that ERK-mediated events were required for binding HuR to the p21 mRNA and preventing its decay. RNA interference-based knockdown of HuR abundance further served to demonstrate the contribution of HuR-mediated p21 mRNA stabilization toward enhancing p21 expression after PGA2 treatment. Collectively, our results indicate that PGA2 stabilizes the p21 mRNA through an ERK-independent increase in cytoplasmic HuR levels and an ERK-dependent association of HuR with the p21 mRNA.

MeSH terms

  • 3' Untranslated Regions
  • Antigens, Surface / chemistry*
  • Biotin / chemistry
  • Blotting, Northern
  • Blotting, Western
  • Calcium / metabolism
  • Cell Cycle Proteins / chemistry
  • Cell Cycle Proteins / metabolism*
  • Cell Line, Tumor
  • Cell Nucleus / metabolism
  • Cell Proliferation
  • Cross-Linking Reagents / pharmacology
  • Cyclin-Dependent Kinase Inhibitor p21
  • Cytoplasm / metabolism
  • DNA, Complementary / metabolism
  • Dactinomycin / pharmacology
  • Dose-Response Relationship, Drug
  • ELAV Proteins
  • ELAV-Like Protein 1
  • Enzyme Inhibitors / pharmacology
  • Humans
  • Immunoprecipitation
  • Kinetics
  • Mitogen-Activated Protein Kinase 3 / metabolism*
  • Prostaglandins A / metabolism*
  • Protein Binding
  • RNA, Messenger / metabolism
  • RNA-Binding Proteins / chemistry*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction
  • Time Factors
  • Transfection
  • Ultraviolet Rays

Substances

  • 3' Untranslated Regions
  • Antigens, Surface
  • CDKN1A protein, human
  • Cell Cycle Proteins
  • Cross-Linking Reagents
  • Cyclin-Dependent Kinase Inhibitor p21
  • DNA, Complementary
  • ELAV Proteins
  • ELAV-Like Protein 1
  • ELAVL1 protein, human
  • Enzyme Inhibitors
  • Prostaglandins A
  • RNA, Messenger
  • RNA-Binding Proteins
  • Dactinomycin
  • Biotin
  • Mitogen-Activated Protein Kinase 3
  • prostaglandin A2
  • Calcium