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KMID : 0620919990310020108
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Experimental & Molecular Medicine
1999 Volume.31 No. 2 p.108 ~ p.114
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Characterization of regulatory elements on the promoter region of human ATP-citrate lyase
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Sahng Wook Park/Young Ah Moon
Kyung Sup Kim/Un Hyung Cho/Do Jun Yoon/Sahng Wook Park
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Abstract
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ATP-citrate lyase (ACL), an enzyme catalyzing the first step in biosynthesis of fatty acids, is induced during the lipogenesis and cholesterologenesis. We demonstrate that the region -213 to -128 of human ACL promoter is responsible for conferring glucose-mediated transcription. This region in the ACL promoter contains Sp1 binding sites determined by DNase I foot-printing assay. Gel retardation assay using oligonucleotides from -179 to -141 and -140 to -110 showed two specific DNA-protein complexes postulated to be formed by transcription factor Sp1. Competition gel shift and supershift assays have confirmed that these DNA-protein complexes were the result of induced Sp1 as well as another Sp1-related proteins. Western blot analysis also demonstrated that transcription factor Sp1 was slightly increased in the nuclear proteins extracted from Alexander cells following supplementation of glucose. In addition, expression of 110 kDa protein reacting with antibody against Sp3 was dramatically increased by glucose supplementation, while isoforms of Sp3, about 80 kDa in size was decreased in its amounts. Our results suggest that changes in the expression of Sp1 family proteins play an important role in activation of the ACL promoter by glucose.
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KEYWORD
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ATP-citrate lyase, promoter, Sp1, Sp3, lipogenesis, glucose,
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