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J Biol Chem, Vol. 275, Issue 18, 13771-13779, May 5, 2000

Post-transcriptional Regulation of Urokinase mRNA
IDENTIFICATION OF A NOVEL UROKINASE mRNA-BINDING PROTEIN IN HUMAN LUNG EPITHELIAL CELLS IN VITRO^*

Sreerama Shetty and Steven Idell

From the Department of Specialty Care Services, The University of Texas Health Center, Tyler, Texas 75708

We sought to determine if urokinase expression is regulated at the post-transcriptional level in cultured lung epithelial cells. We also sought to determine if differences in urokinase expression by cultured human lung carcinoma and non-malignant lung epithelial subtypes were attributable to post-transcriptional regulatory mechanisms. Urokinase was expressed by phenotypically diverse lung carcinoma cell lines as well as non-malignant small airway epithelial cells and bronchial epithelial cells. Using gel mobility shift and UV cross-linking assays, we identified a 30-kDa urokinase mRNA-binding protein that selectively bound to a 66-nucleotide protein-binding fragment of urokinase mRNA. The urokinase mRNA-binding protein is found in the cytosolic but not nuclear extracts of non-malignant lung epithelial cells; whereas, it is found in the nuclear but not cytosolic extracts of selected malignant carcinoma-derived cells that express relatively large amounts of urokinase. Chimeric -globin/urokinase cDNA containing the urokinase mRNA-binding protein binding sequence destabilized otherwise stable -globin mRNA. Our results demonstrate that urokinase gene expression in lung epithelial and lung carcinoma-derived cells is regulated at the post-transcriptional level. The mechanism involves an interaction between a 66-nucleotide sequence of the urokinase mRNA 3'-untranslated region with a newly recognized urokinase mRNA-binding protein to regulate urokinase mRNA stability.

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