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Originally published In Press as doi:10.1074/jbc.M010627200 on January 26, 2001

J. Biol. Chem., Vol. 276, Issue 17, 13675-13684, April 27, 2001
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Plasminogen Activator Inhibitor Type 2 Contains mRNA Instability Elements within Exon 4 of the Coding Region
SEQUENCE HOMOLOGY TO CODING REGION INSTABILITY DETERMINANTS IN OTHER mRNAs*

Marcus J. Tierney and Robert L. MedcalfDagger

From the Department of Medicine, Monash University, Box Hill Hospital, Box Hill 3128, Victoria, Australia

Plasminogen activator inhibitor type 2 (PAI-2) is a serine protease inhibitor that inhibits urokinase. Constitutive and regulated PAI-2 gene expression involves post-transcriptional events, and an AU-rich mRNA instability motif within the 3'-untranslated region of PAI-2 mRNA is required for this process (Maurer, F., Tierney, M., and Medcalf, R. L. (1999) Nucleic Acids Res. 27, 1664-1673). Here we show that instability determinants are present within various exons of the PAI-2 coding region, most notably within exon 4. Deletion of exon 4 from the full-length PAI-2 cDNA results in a doubling in the half-life of PAI-2 mRNA, whereas a 28-nucleotide region within exon 4 contains binding sites for cytoplasmic proteins. Inducible stabilization of PAI-2 mRNA in HT-1080 cells treated with phorbol ester and tumor necrosis factor does not alter the binding of proteins to the exon 4 instability determinant, but resulted in a transient increase in the binding of factors to the AU-rich RNA instability element. Hence, PAI-2 mRNA stability is influenced by elements located within both the coding region and the 3'-untranslated region and that cytoplasmic mRNA binding factors may influence steady state and inducible PAI-2 mRNA expression. Finally a 10-nucleotide region flanking the exon 4 protein-binding site is homologous to instability elements within five other transcripts, suggesting that a common coding region determinant may exist.

* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Dept. of Medicine, Monash University, Box Hill Hospital, Box Hill 3128, Victoria, Australia. Tel.: 61 3 9895 0318; Fax: 61 3 9895 0332; E-mail: robert. medcalf@med.monash.edu.au.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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