Chemical inhibitors destabilize HuR binding to the AU-rich element of TNF-alpha mRNA

Exp Mol Med. 2009 Nov 30;41(11):824-31. doi: 10.3858/emm.2009.41.11.088.

Abstract

Hu protein R (HuR) binds to the AU-rich element (ARE) in the 3UTR to stabilize TNF-alpha mRNA. Here, we identified chemical inhibitors of the interaction between HuR and the ARE of TNF-alpha mRNA using RNA electrophoretic mobility gel shift assay (EMSA) and filter binding assay. Of 179 chemicals screened, we identified three with a half-maximal inhibitory concentration (IC(50)) below 10 microM. The IC(50) of quercetin, b-40, and b-41 were 1.4, 0.38, and 6.21 microM, respectively, for binding of HuR protein to TNF-alpha mRNA. Quercetin and b-40 did not inhibit binding of tristetraprolin to the ARE of TNF-alpha mRNA. When LPS-treated RAW264.7 cells were treated with quercetin and b-40, we observed decreased stability of TNF-alpha mRNA and decreased levels of secreted TNF-alpha. From these results, we could find inhibitors for the TNF-alpha mRNA stability, which might be used advantageously for both the study for post-transcriptional regulation and the discovery of new anti-inflammation drugs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions*
  • Animals
  • Anti-Inflammatory Agents / pharmacology*
  • Antigens, Surface / metabolism
  • Antioxidants / pharmacology
  • Cell Line
  • Dose-Response Relationship, Drug
  • Drug Evaluation, Preclinical
  • ELAV Proteins
  • ELAV-Like Protein 1
  • Mice
  • Protein Binding / drug effects
  • Quercetin / pharmacology*
  • RNA Stability / drug effects*
  • RNA-Binding Proteins / antagonists & inhibitors*
  • RNA-Binding Proteins / metabolism
  • Tumor Necrosis Factor-alpha / biosynthesis*

Substances

  • 3' Untranslated Regions
  • Anti-Inflammatory Agents
  • Antigens, Surface
  • Antioxidants
  • ELAV Proteins
  • ELAV-Like Protein 1
  • ELAVL1 protein, human
  • RNA-Binding Proteins
  • Tumor Necrosis Factor-alpha
  • Quercetin