A 40-bp RNA element that mediates stabilization of vascular endothelial growth factor mRNA by HuR.
VEGF is a critical mediator of hypoxia-induced angiogenesis in numerous physiological and pathophysiological conditions. The hypoxic induction of VEGF is due in large part to an increase in the stability of its mRNA. We recently demonstrated that the stabilization of VEGF mRNA by hypoxia is dependent upon the RNA-binding protein HuR. This report describes the identification of a 40-bp functional HuR binding site in the VEGF mRNA 3'-untranslated region. This element can confer HuR-mediated stabilization of a heterologous gene in vitro and in vivo. Furthermore, the element is sufficient to confer an increase in the hypoxic induction of a heterologous gene. Deletion of the HuR binding site within this 40-bp element as mapped by RNase T1 and lead footprinting uncouples a stabilizing sequence from a destabilizing sequence, thus providing a novel RNA-protein regulatory model that might be exploited to manipulate VEGF expression and hypoxia-induced angiogenesis.
Goldberg-Cohen
Ilana
I
Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Post Office Box 9649, Haifa 31096, Israel.
Furneauxb
Henry
H
Levy
Andrew P
AP
D020413:3' Untranslated Regions; D000954:Antigens, Surface; D001665:Binding Sites; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D051959:ELAV Proteins; D000067780:ELAV-Like Protein 1; D018274:Electroporation; D016228:Endothelial Growth Factors / Q000378:metabolism*; D017930:Genes, Reporter; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D000860:Hypoxia; D008156:Luciferases / Q000378:metabolism; D008222:Lymphokines / Q000378:metabolism*; D009154:Mutation; D018919:Neovascularization, Physiologic; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism; D009418:S100 Proteins / Q000378:metabolism; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D020413:3' Untranslated Regions; D000954:Antigens, Surface; D051959:ELAV Proteins; D000067780:ELAV-Like Protein 1; C098770:ELAVL1 protein, human; D016228:Endothelial Growth Factors; D008222:Lymphokines; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D009418:S100 Proteins; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D012313:RNA; D008156:Luciferases; D005982:Glutathione Transferase