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Vol. 9, Issue 2, 469-481, February 1998
Untranslated Region
Mediating Hypoxia-induced mRNA Stability
and
Departments of
*Pathology,
Hypoxia is a prominent feature of malignant tumors that are
characterized by angiogenesis and vascular hyperpermeability. Vascular
permeability factor/vascular endothelial growth factor (VPF/VEGF) has
been shown to be up-regulated in the vicinity of necrotic tumor areas,
and hypoxia potently induces VPF/VEGF expression in several tumor cell
lines in vitro. Here we report that hypoxia-induced VPF/VEGF expression
is mediated by increased transcription and mRNA stability in human M21
melanoma cells. RNA-binding/electrophoretic mobility shift assays
identified a single 125-bp AU-rich element in the 3Medicine, and
§Dermatology, Beth Israel Deaconess Medical Center and
Harvard Medical School, Boston, Massachusetts 02215
untranslated
region that formed hypoxia-inducible RNA-protein complexes.
Hypoxia-induced expression of chimeric luciferase reporter constructs
containing this 125-bp AU-rich hypoxia stability region were
significantly higher than constructs containing an adjacent 3
untranslated region element without RNA-binding activity. Using UV-cross-linking studies, we have identified a series of
hypoxia-induced proteins of 90/88 kDa, 72 kDa, 60 kDa, 56 kDa, and 46 kDa that bound to the hypoxia stability region element. The 90/88-kDa
and 60-kDa species were specifically competed by excess hypoxia
stability region RNA. Thus, increased VPF/VEGF mRNA stability induced
by hypoxia is mediated, at least in part, by specific interactions between a defined mRNA stability sequence in the 3 untranslated region
and distinct mRNA-binding proteins in human tumor cells.
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