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Inherent instability of plasminogen activator inhibitor type 2 mRNA is regulated by tristetraprolin

Hong Yu1, Stan Stasinopoulos, Peter Leedman

  • 1Department of Medicine, Monash University, Box Hill Hospital, Victoria, Australia.

The Journal of Biological Chemistry
|February 13, 2003
PubMed

Insights

Tristetraprolin (TTP) binds to the PAI-2 mRNA AU-rich element (ARE), destabilizing PAI-2 mRNA and reducing protein levels. This post-transcriptional regulation occurs even at very low TTP concentrations.

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • RNA Metabolism

Background:

  • Plasminogen activator inhibitor type 2 (PAI-2) gene expression is regulated post-transcriptionally.
  • PAI-2 mRNA contains instability elements, including an AU-rich element (ARE) in the 3'-untranslated region (UTR).
  • The HuR protein is known to bind the PAI-2 ARE and affect mRNA stability.

Purpose of the Study:

  • To identify novel proteins that bind to the PAI-2 ARE.
  • To investigate the role of identified proteins in PAI-2 mRNA regulation.
  • To characterize the functional impact of protein binding on PAI-2 mRNA stability and protein expression.

Main Methods:

  • Yeast three-hybrid system screening of a human leukocyte cDNA library.
  • UV cross-linking and immunoprecipitation (IP) assays.
  • Co-transfection experiments in HEK293 cells with PAI-2 and candidate protein expression plasmids.
  • Analysis of reporter transcript stability.

Main Results:

  • Tristetraprolin (TTP) was identified as a PAI-2 ARE-binding protein.
  • TTP directly associates with the PAI-2 ARE in vitro and in vivo.
  • TTP expression accelerates PAI-2 mRNA decay and reduces PAI-2 protein levels in a 3'-UTR-dependent manner.
  • The PAI-2 ARE confers TTP-dependent instability to a reporter mRNA.
  • PAI-2 mRNA decay is sensitive to TTP, occurring at sub-detection levels.

Conclusions:

  • TTP is a functional PAI-2 ARE-binding protein.
  • TTP mediates post-transcriptional silencing of the PAI-2 gene by promoting mRNA decay.
  • This study elucidates a novel mechanism for PAI-2 gene regulation involving TTP-mediated mRNA instability.

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