Common Proteins Bind mRNAs Encoding Erythropoietin, Tyrosine Hydroxylase, and Vascular Endothelial Growth Factor

Abstract

The hypoxia-inducible genes erythropoietin (Epo), tyrosine hydroxylase (TH), and vascular endothelial growth factor (VEGF) are regulated post-transcriptionally by proteins binding to specific regions located in the 3′ untranslated region (UTR) of their mRNAs. To determine whethertrans-factors binding to this region in all three of these RNAs are similar, we generated riboprobes containing the 3′ UTR of erythropoietin, tyrosine hydroxylase, and vascular endothelial growth factor mRNA and assayed them by electrophoretic mobility shift assay (EMSA) and UV cross-linking experiments. Each riboprobe formed similar shifted protein complexes using human hepatoma cell (Hep3B) cytoplasmic lysates in the EMSA. Hep3B proteins bound to each probe could be cross-competed by the specific unlabeled Epo, TH, or VEGF riboprobes. By contrast, a non-specific 3′ UTR riboprobe did not compete for binding with the Epo, TH, or VEGF RNA shifted protein complexes. UV cross-linking studies revealed proteins of similar molecular weights for the Epo, TH, and VEGF RNA shifted protein complexes. Taken together, these results suggest a common post-transcriptional regulatory mechanism for hypoxia-inducible genes.