Expression of GDH gene and regulation mechanism of the GDH gene function by HIF-1α gene in sea urchin(Strongylocentrotus intermedius)
In order to elucidate the molecular mechanism of GDH gene(glutamate dehydrogenase,GDH)expres-sion and function regulated by the transcription factor HIF-1α gene in sea urchins(Strongylocentrotus intermedius),the promoter sequence of GDH gene and its structural characteristics were cloned and analyze by genome walking,luciferase reporter assay and RNA interference techniques and the binding site and regulatory relationship between HIF-1α gene and the promoter region of GDH gene were verified in S.intermedius.The effect of HIF-1α gene regu-lation on the expression of GDH gene on the energy production of coelomocytes and gonads of S.intermedius was in-vestigated.The results showed that the total length of GDH gene promoter was found to be 1 067 bp,containing 12 promoter elements,14 different transcription factor binding sites and 1 CpG island,with core promoter region ran-ging from-984 to-363 bp.There were two binding sites between HIF-1α gene and GDH gene promoter region(-683 to-649 bp;-280 to-264 bp),with a positive regulatory relationship between the two.The relative ex-pression of GDH(mRNA and protein levels)and the activity of relative total GDH in coelomocytes and gonads were shown to be a downward trend,accompanied by a significant decrease in the relative content of ATP in S.interme-dius exposed to inhibition of HIF-1α gene expression.These findings suggest that HIF-1α gene in sea urchin can af-fect the expression of GDH gene by positively regulating its transcriptional activity,and ultimately affect the ATP level in different tissues of the sea urchin.
Strongylocentrotus intermediushypoxia inducible factor-1αGDH gene promotertranscriptional regu-lationenergy production
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维普
