Cholesteryl ester transfer protein gene expression is not specifically regulated by CCAAT/enhancer-binding protein in HepG2-cells.
Cholesteryl ester transfer protein (CETP) mediates the exchange of neutral lipids among plasma lipoproteins and is expressed predominantly in liver and intestine. In band shift assays employing nuclear extracts of HepG2 cells we identified C/EBPbeta as the predominant C/EBP isoform involved in binding to the C/EBP consensus sequence within the 5' upstream region of the CETP gene. This was demonstrated by supershift experiments using antibodies specific for C/EBPalpha, C/EBPbeta and C/EBPdelta and an oligonucleotide containing a single point mutation (CAAT-->CTAT) in this site. Expression of a CETP promoter-fragment/luciferase construct in transiently transfected HepG2 and CaCo-2 cells and enhancement of promoter activity by co-transfection with human C/EBPalpha in HepG2 cells could be influenced neither by the mutation in the consensus sequence nor by elimination of this site together with a second potential binding site for C/EBP. Furthermore, transfection of HepG2 with human C/EBPalpha did not influence the synthesis of CETP by these cells. Our results indicate that the expression of C/EBP in HepG2 cells is not able (1) to influence specifically the expression of a transfected CETP promoter dependent reporter through binding to C/EBP sites in the promoter region and (2) to significantly enhance expression of the endogenous CETP gene.
Ritsch
A
A
Department of Medicine, University of Innsbruck, Austria.
Doppler
W
W
Pfeifhofer
C
C
Sandhofer
A
A
Bodner
J
J
Patsch
J R
JR
D001053:Apolipoproteins / Q000097:blood / Q000235:genetics*; D001483:Base Sequence; D022762:CCAAT-Enhancer-Binding Proteins; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics*; D002460:Cell Line; D053480:Cholesterol Ester Transfer Proteins; D004268:DNA-Binding Proteins / Q000032:analysis / Q000378:metabolism / Q000502:physiology*; D004742:Enhancer Elements, Genetic; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D005786:Gene Expression Regulation / Q000502:physiology*; D006023:Glycoproteins; D006801:Humans; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008156:Luciferases / Q000032:analysis / Q000378:metabolism; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D001616:beta-Galactosidase / Q000032:analysis / Q000378:metabolism
D001053:Apolipoproteins; D022762:CCAAT-Enhancer-Binding Proteins; C506642:CETP protein, human; D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D004268:DNA-Binding Proteins; D006023:Glycoproteins; D009687:Nuclear Proteins; D008156:Luciferases; D001616:beta-Galactosidase