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Articles in PresS, published online ahead of print May 23, 2002
Am J Physiol Heart Circ Physiol, 10.1152/ajpheart.00813.2001
Submitted on September 17, 2001
Accepted on May 20, 2002
1 Medicine/Physiology, University of California, San Diego, La Jolla, CA, USA
* To whom correspondence should be addressed. E-mail: ktang{at}ucsd.edu.
Hypoxic exercise increases VEGF expression and the formation of new capillaries. In addition to HIF 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 post-transcriptional regulator of VEGF gene expression. Here, we report that in rat skeletal muscle (gastrocnemius): (1) HuR binds to a known regulatory sequence located in the VEGF mRNA 3'-UTR (1631-1678 bp); (2) HuR specifically binds in to the A/U rich element AUUUUA (1665-1670 bp) and an additional A/U rich region containing the consensus sequence UUUUUUA (1658-1664 bp); (3) binding of HuR to VEGF mRNA is seen already 5 min after acute ischemia, remaining elevated throughout a 60-minute ischemic period; (4) a second inducible HuR-VEGF mRNA binding factor is evident 30 and 60 minutes post-ischemia; and (5) VEGF mRNA and protein levels are increased 20 min and 30 min, respectively, after acute ischemia. These findings suggest that acute ischemia induces a rapid binding of HuR to VEGF mRNA 3'UTR. In skeletal muscle, this specific protein-RNA interaction may be an important post-transcriptional regulatory mechanism for increasing VEGF expression in response to hypoxia or acute ischemia.
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