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Division of Physiology, Department of Medicine, University of California, La Jolla, California 92093
Hypoxic exercise increases VEGF expression and the formation of new capillaries. In addition to hypoxia-inducible factor regulation at the transcriptional level, VEGF message stabilization is also a key regulatory step for VEGF expression. In vitro experiments have identified Hu protein R (HuR) as a potential posttranscriptional regulator of VEGF gene expression. Here, we report that in rat skeletal muscle (gastrocnemius muscle), 1) HuR binds to a known regulatory sequence located in the VEGF mRNA 3'-untranslated region (1,631-1,678 bp); 2) HuR specifically binds to the A/U-rich element AUUUUA (1,665-1,670 bp) and an additional A/U-rich region containing the consensus sequence UUUUUUA (1,658-1,664 bp); 3) binding of HuR to VEGF mRNA is seen already 5 min after acute ischemia, remaining elevated throughout a 60-min ischemic period; 4) a second inducible HuR-VEGF mRNA binding factor is evident 30 and 60 min postischemia; and 5) VEGF mRNA and protein levels are increased 20 and 30 min, respectively, after acute ischemia. These findings suggest that acute ischemia induces a rapid binding of HuR to the VEGF mRNA 3'-untranslated region. In skeletal muscle, this specific protein-RNA interaction may be an important posttranscriptional regulatory mechanism for increasing VEGF expression in response to hypoxia or acute ischemia.
angiogenesis; acute ischemia; hypoxia
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