Inherent instability of plasminogen activator inhibitor type 2 mRNA is regulated by tristetraprolin
| Autor: | Hong Yu, Peter J. Leedman, Robert L. Medcalf, Stan Stasinopoulos |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Untranslated region
DNA Complementary Transcription Genetic Ultraviolet Rays Tristetraprolin Blotting Western RNA-binding protein Biology Transfection Biochemistry Immediate early protein Cell Line ELAV-Like Protein 1 Immediate-Early Proteins Fungal Proteins Two-Hybrid System Techniques Plasminogen Activator Inhibitor 2 Coding region Humans RNA Messenger RNA Processing Post-Transcriptional Molecular Biology 3' Untranslated Regions Gene Library Messenger RNA Dose-Response Relationship Drug Models Genetic Three prime untranslated region cDNA library RNA-Binding Proteins Cell Biology Blotting Northern Molecular biology Precipitin Tests Globins DNA-Binding Proteins Cross-Linking Reagents ELAV Proteins Antigens Surface Protein Binding |
| Zdroj: | The Journal of biological chemistry. 278(16) |
| ISSN: | 0021-9258 |
| Popis: | Plasminogen activator inhibitor type 2 (PAI-2) is a serine protease inhibitor that is subject to regulation at the post-transcriptional level. At least two mRNA instability elements reside within the PAI-2 transcript; one in the coding region and another within the 3'-untranslated region (UTR). For the latter, a functional AU-rich motif (ARE) has been identified that provides a binding site for a number of cellular proteins, including the mRNA stability protein, HuR. In this study, we used the yeast three-hybrid system to screen a human leukocyte cDNA library to identify other proteins that associate with the PAI-2 ARE. This screen identified tristetraprolin (TTP) as a PAI-2 mRNA ARE-binding protein. UV cross-linking and immunoprecipitation experiments showed that TTP expressed in HEK293 cells could associate with the PAI-2 ARE in vitro. Co-transfection of plasmids expressing TTP and PAI-2 in HEK293 cells resulted in an increase in the decay rate of PAI-2 mRNA and loss of PAI-2 protein in a process that was dependent upon the PAI-2 3'-UTR. The 29-nt PAI-2 AU-rich element alone was also capable of conferring TTP-dependent mRNA instability to a reporter transcript. The extent of PAI-2 mRNA stability was remarkably sensitive to TTP since TTP-dependent PAI-2 mRNA decay occurred at TTP levels that were below Western blot detection limits. This study identifies TTP as a functional PAI-2 ARE-binding protein that modulates the post-transcriptional regulation of the PAI-2 gene. |
| Databáze: | OpenAIRE |
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