Regulatory elements in the promoter region of the rat gene encoding the acyl-CoA-binding protein.
Acyl-CoA-binding protein (ACBP) is an ubiquitously expressed 10-kDa protein which is present in high amounts in cells involved in solute transport or secretion. Rat ACBP is encoded by a gene containing the typical hallmarks of a housekeeping gene. Analysis of the promoter region of the rat ACBP gene by electrophoretic mobility shift assay (EMSA) revealed specific binding of proteins from rat liver nuclear extracts to potential recognition sequences of NF-1/CTF, Sp1, AP-1, C/EBP and HNF-3. In addition, specific binding to a DR-1 type element was observed. By using in vitro translated peroxisome proliferator activated receptors (PPAR) and a retinoid X receptor alpha (RXRalpha), we demonstrated that this DR-1 element was capable of binding PPARalpha/RXRalpha, PPARdelta/RXRalpha and PPARgamma2/RXRalpha heterodimers. The PPARgamma2/RXRalpha heterodimer appeared to have the highest affinity for the ACBP DR-1 element. Addition of peroxisome proliferators (PP) to H4IIEC3 rat hepatoma cells led to an increase in the ACBP mRNA level, indicating that the DR-1 element could be a functional peroxisome proliferator responsive element (PPRE). Analysis of the ACBP promoter by transient transfection showed that deletion of the region containing the DR-1 element reduced transcriptional activity, and further indicated that three AP-2 sites and one NF-1/CTF site in the proximal promoter are of importance for basal promoter activity.
Elholm
M
M
Department of Molecular Biology, University of Odense, Denmark.
Bjerking
G
G
Knudsen
J
J
Kristiansen
K
K
Mandrup
S
S
D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000235:genetics*; D026261:Diazepam Binding Inhibitor; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression / Q000187:drug effects; D010750:Phosphoproteins / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011743:Pyrimidines / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
D002352:Carrier Proteins; D026261:Diazepam Binding Inhibitor; D010750:Phosphoproteins; D011743:Pyrimidines; D014157:Transcription Factors; C075995:down-regulator of transcription 1; C006253:pirinixic acid