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A more recent version of this article appeared on May 1, 2003

Papers In Press, published online ahead of print February 16, 2003
J. Lipid Res., doi:10.1194/jlr.M200423-JLR200
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Submitted on October 25, 2002
Revised on January 30, 2003
Accepted on February 11, 2003

A CPF site and ALU repeat in the distal promoter region are implicated in regulation of human CETP gene expression

Wilfried Le Goff, Maryse Guérin, M. John Chapman, and Joëlle Thillet

INSERM U551, Hôpital de la Pitié, Paris 75651

Corresponding Author: chapman{at}chups.jussieu.fr

The cholesteryl ester transfer protein (CETP) plays a key role in reverse cholesterol transport in mediating the transfer of cholesteryl ester (CE) from high density lipoproteins (HDL) to atherogenic ApoB-containing lipoproteins (VLDL, IDL and LDL) and thereby facilitating CE uptake by the liver. Variation in plasma CETP mass in both normolipidemic and in dyslipidemic individuals may reflect differences in CETP gene expression. As the 5' flanking sequence up to 3.4 kb of the human CETP gene contributes to transcriptional activity and tissue-specific gene expression, we evaluated the role of the distal promoter region in the modulation of CETP gene expression. Deletional analysis in transient transfections in HepG2 cells using luciferase reporter constructs containing a variable length of the human CETP promoter (from -1012 bp up to -3242 bp) permitted the identification of an activator (-1012/-1398) and a repressor (-2146/-2680) promoter region. The use of chimeric constructs revealed that an Alu repeat (-2153/-2414) accounted for repression observed in the region from -2146 to -2680. A binding site for the orphan nuclear receptor CPF was identified at position -1042, which partially accounted for the activation mediated by the enhancing region in HepG2 cells; in contrast, this site was inactive in HEK293 cells. Co-transfection of LRH, the mouse homologue of CPF, in HEK293 cells, indicated that the -1042 CPF site is sufficient to induce CPF-mediated activation of CETP promoter activity to a similar degree as the previously described -75 CPF site. Taken together, our results indicate that the distal promoter region is a major component of the modulation of human CETP promoter activity and that it may contribute to the liver-specific expression of the CETP gene.


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