Cloning of the human cholesteryl ester hydrolase promoter: identification of functional peroxisomal proliferator-activated receptor responsive elements

Biochem Biophys Res Commun. 2001 Jun 22;284(4):1065-70. doi: 10.1006/bbrc.2001.5078.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Pairing
  • Base Sequence
  • Binding Sites
  • COS Cells
  • Chlorocebus aethiops
  • Cloning, Molecular / methods
  • Consensus Sequence
  • DNA Primers
  • DNA-Binding Proteins / metabolism
  • Genes, Reporter
  • Genomic Library
  • Humans
  • Luciferases / genetics
  • Macrophages / enzymology
  • Molecular Sequence Data
  • Promoter Regions, Genetic*
  • Receptors, Cytoplasmic and Nuclear / metabolism*
  • Sp1 Transcription Factor / metabolism
  • Sterol Esterase / genetics*
  • Transcription Factors / metabolism*
  • Transfection

Substances

  • DNA Primers
  • DNA-Binding Proteins
  • Receptors, Cytoplasmic and Nuclear
  • Sp1 Transcription Factor
  • Transcription Factors
  • Luciferases
  • Sterol Esterase