Cloning of the Human Phospholipase C-γ1 Promoter and Identification of a DR6-type Vitamin D-responsive Element
作者： Zhongjian Xie , Daniel D. Bikle
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摘要: The 5′-flanking region of the human phospholipase C-γ1 gene was isolated from a P1 genomic DNA library. S1-nuclease mapping and primer extension analysis revealed that there is single transcriptional start site located at 135 bases upstream translation codon in gene. sequence showed around very GC-rich has no TATA box. fragment +135 to −877 subcloned into luciferase reporter vector. chimeric produced high level activity responded 1,25-(OH)2D3 transiently transfected keratinocytes. Deletion mutation studies demonstrated vitamin D-responsive element contains motif arranged as two direct repeats separated by 6 (DR6), AGGTCAgaccacTGGACA, between −786 −803 base pairs. Incubation oligonucleotide containing DR6 with keratinocyte nuclear extracts specific protein-DNA complex shifted higher molecular weight form upon addition an antibody receptor. Therefore, confers promoter DR6-type mediates, least part, enhanced expression this keratinocytes 1,25-(OH)2D3.
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