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Papers In Press, published online ahead of print February 11, 2002
Institut for clinical chemistry and laboratory medicine, Universitiy of Regensburg, Regensburg 93053
Corresponding Author: gerd.schmitz{at}klinik.uni-regensburg.de
The ATP-binding cassette transporter A1 (ABCA1) shows a differentiation-, cAMP- and sterol dependent upregulation in human monocytes. As part of an ongoing study, we investigated the proximal promoter regions which are highly conserved between the human and murine ABCA1 genes. Using reporter gene assays we show here that a TATA box 24 bp upstream of the transcription initiation site is essential for promoter activity in RAW264.7 and HepG2 cells, while further enhancement of transcriptional activity is mediated by the -175 bp promoter region. Gelshift assays revealed in vitro binding of Sp1 to a -91 GnC motif as well as binding of Sp1 and Sp3 to a -157 GnC promoter region. In co-transfection experiments using Drosophila S2 cells we demonstrate that Sp3 competes with Sp1 for binding to the -157 GnC motif and acts as a repressor. On the other hand, overexpression of Sp1 increased ABCA1 mRNA expression in HeLa cells and enhanced cellular cholesterol and phospholipid efflux in RAW246.7 macrophages. Besides the functional importance for basal gene expression we have identified that the core promoter region (-175 to +229) is also responsible for the induction of ABCA1 by the cytokine Oncostatin M resulting in a rapid increase in ABCA1 mRNA levels in HepG2 cells.
J. Biol. Chem, 10.1074/jbc.M110270200
Submitted on October 25, 2001
Revised on January 23, 2002
Accepted on February 9, 2002
Identification of sterol-independent regulatory elements in the human atp-binding cassette transporter A1 (ABCA1) promoter: role of Sp1/3, E-Box binding factors and an oncostatin m responsive element
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