Hypoxia-inducible protein binding to vascular endothelial growth factor mRNA and its modulation by the von Hippel-Lindau protein.
Hypoxia induces an increase in the stability of the mRNA encoding vascular endothelial growth factor (VEGF). We have previously demonstrated that a 500-base region of the 3'-untranslated region of VEGF mRNA that is critical for stabilization of VEGF mRNA in an in vitro degradation assay forms a RNA-protein complex in a hypoxia-inducible fashion. We report here the identification of three adenylate-uridylate-rich RNA elements within this region that form an identical or closely related hypoxia-inducible RNA-protein complex. This complex is constitutively elevated in a tumor cell line lacking the wild type von Hippel-Lindau tumor suppressor gene and in which VEGF mRNA is constitutively stabilized. Furthermore, the glucose transporter-1 mRNA, which is also stabilized by hypoxia, forms a hypoxia-inducible RNA-protein complex with similar sequence and protein binding characteristics to that described for VEGF mRNA. Finally, RNA affinity purification and UV cross-linking were used to identify three proteins of 32, 28, and 17 kDa that are derived from this hypoxia-inducible RNA-protein complex.
Levy
A P
AP
Whitaker Cardiovascular Institute, Evans Department of Medicine, Boston University School of Medicine, Boston, Massachusetts 02118, USA.
Levy
N S
NS
Goldberg
M A
MA
D000225:Adenine; D000818:Animals; D001482:Base Composition; D001483:Base Sequence; D002417:Cattle; D015687:Cell Hypoxia; D002460:Cell Line; D017931:DNA Primers; D016228:Endothelial Growth Factors / Q000096:biosynthesis* / Q000235:genetics; D017353:Gene Deletion; D016147:Genes, Tumor Suppressor; D051272:Glucose Transporter Type 1; D006801:Humans; D008025:Ligases; D008222:Lymphokines / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins / Q000096:biosynthesis*; D016376:Oligonucleotides, Antisense; D011485:Protein Binding; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D051381:Rats; D013698:Templates, Genetic; D014407:Tumor Cells, Cultured; D025521:Tumor Suppressor Proteins; D044767:Ubiquitin-Protein Ligases; D014466:Ultraviolet Rays; D014498:Uracil; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D051794:Von Hippel-Lindau Tumor Suppressor Protein
D017931:DNA Primers; D016228:Endothelial Growth Factors; D051272:Glucose Transporter Type 1; D008222:Lymphokines; D009004:Monosaccharide Transport Proteins; D016376:Oligonucleotides, Antisense; D011506:Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C495677:SLC2A1 protein, human; C495679:Slc2a1 protein, rat; D025521:Tumor Suppressor Proteins; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D014498:Uracil; D044767:Ubiquitin-Protein Ligases; D051794:Von Hippel-Lindau Tumor Suppressor Protein; D008025:Ligases; C497446:VHL protein, human; D000225:Adenine