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Molecular and Cellular Biology, October 1999, p. 6471-6478, Vol. 19, No. 10
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Identification of a Novel Dexamethasone-Sensitive RNA-Destabilizing Region on Rat Monocyte Chemoattractant Protein 1 mRNA

Michael Poon,* Bin Liu, and Mark B. Taubman

The Zena and Michael A. Wiener Cardiovascular Institute and Department of Medicine, Mount Sinai School of Medicine, New York, New York

Received 2 April 1999/Returned for modification 7 May 1999/Accepted 9 July 1999

Glucocorticoids are potent anti-inflammatory agents widely used in the treatment of human disease. We have previously shown that the inflammatory cytokine monocyte chemoattractant protein 1 (MCP-1) is regulated posttranscriptionally by glucocorticoids in arterial smooth muscle cells (SMC). To elucidate the mechanism mediating this effect, in vitro-transcribed radiolabeled MCP-1 mRNA was incubated with cytoplasmic extracts from SMC and analyzed by gel electrophoresis. Extracts from SMC treated with platelet-derived growth factor (PDGF) did not degrade the transcripts for up to 3 h. In contrast, extracts from cells treated with 1 µM dexamethasone (Dex) alone or in combination with PDGF degraded the probe with a half-life of approx 15 min. Dex had maximal effect at concentrations above 0.01 µM and was effective on both rat and human MCP-1 transcripts. By deletion analysis, the Dex-sensitive region of the MCP-1 mRNA was localized to the initial 224 nucleotides (nt) at the 5' end and did not involve an AU-rich sequence in the 3' untranslated end. The 224-nt region conferred Dex sensitivity to heterologous mRNA. These studies provide new insights into the molecular mechanisms underlying the effect of glucocorticoids on gene expression.

* Corresponding author. Mailing address: Box 1030, Mount Sinai School of Medicine, One Gustave L. Levy Place, New York, NY 10029. Phone: (212) 241-3913. Fax: (212) 987-3258. E-mail: m_poon{at}smtplink.mssm.edu.

Molecular and Cellular Biology, October 1999, p. 6471-6478, Vol. 19, No. 10
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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