Cloning of the human cholesteryl ester hydrolase promoter: identification of functional peroxisomal proliferator-activated receptor responsive elements.
Cholesteryl ester hydrolase (CEH) is responsible for hydrolysis of stored cholesterol esters in macrophage foam cells and release of free cholesterol for high-density lipoprotein-mediated efflux. PCR-based screening of human genomic libraries with human macrophage CEH specific primers resulted in amplification and cloning of 1.7 kb promoter sequence. Analysis of the sequence revealed a lack of consensus TATA-box but presence of a GC-rich proximal sequence, a CAAT box and several binding sites for the transcription factor Sp1. Three putative response elements for peroxisome proliferator-activated receptor (PPRE) were identified at position -176, -779, and -1316. Down-regulation of promoter activity was observed in the presence of either PPARalpha- or PPARgamma-specific ligands and introduction of a 4-point transverse mutation in the PPRE at -176 completely abolished the effect of PPAR ligands on the promoter activity. Analogous to other genes involved in macrophage cholesterol homeostasis, human CEH may also be regulated by PPAR.
Ghosh
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S
Department of Internal Medicine, Virginia Commonwealth University, Richmond, Virginia 23298-0050, USA. shobha@hsc.vcu.edu
Natarajan
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R
D000595:Amino Acid Sequence; D000818:Animals; D020029:Base Pairing; D001483:Base Sequence; D001665:Binding Sites; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular / Q000379:methods; D016384:Consensus Sequence; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000378:metabolism; D017930:Genes, Reporter; D015698:Genomic Library; D006801:Humans; D008156:Luciferases / Q000235:genetics; D008264:Macrophages / Q000201:enzymology; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D016329:Sp1 Transcription Factor / Q000378:metabolism; D002787:Sterol Esterase / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism*; D014162:Transfection
D017931:DNA Primers; D004268:DNA-Binding Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D016329:Sp1 Transcription Factor; D014157:Transcription Factors; D008156:Luciferases; D002787:Sterol Esterase