Exp Mol Med.  1999 Jun;31(2):108-114.

Characterization of regulatory elements on the promoter region of human ATP-citrate lyase

• Moon YA
• Kim KS
• Cho UH
• Yoon DJ
• Park SW

Abstract

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.

Keyword

ATP-citrate lyase; promoter; Sp1; Sp3; lipogenesis; glucose

MeSH Terms

ATP Citrate (pro-S)-Lyase/metabolism
ATP Citrate (pro-S)-Lyase/genetics*
Binding Sites
Cells, Cultured
Chloramphenicol O-Acetyltransferase/genetics
DNA Footprinting/methods
Deoxyribonuclease I/metabolism
Electrophoresis, Polyacrylamide Gel
Gene Expression Regulation, Enzymologic*
Glucose/pharmacology
Glucose/metabolism*
Human
Immunoblotting
Promoter Regions (Genetics)*
Transcription Factor, Sp1/metabolism*
Transcription, Genetic*
Transfection