| Objective: The aim of this project was to investigate the basic transcriptionalregulation mechanisms of ApoF。Methods: The full length promoter ApoF and a series of truncated promoter regionswere cloned. A luciferase reporter assay revealed the key transcription regulationdomain. Furthermore, TFSEARCH and TESS programs were used to predict potentialtranscription factor binding sites. Site-directed mutagenesis, overexpressiontranscription factors and dominant negative mutants, electrophoretic mobility shiftassay and chromatin immunoprecipitation were used to further confirm binding sites.Results:1.Regions of-198nt to+79nt are crucial for the full activities of ApoFpromoter.2.Identification of ETS and C/EBP binding sites in-198nt to+79nt regionof ApoF promoter.3.ApoF promoter displayed highest activities in liver-derived celllines.4.ETS-1, ETS-2expression clones and their dominant negative mutants wassuccessfully constructed.5.Overexpression of ETS-1and ETS-2enhanceapolipoprotein F promoter activities.Conclusion:1. ApoF is liver tissue-specific gene.2. ApoF gene is regulated by ETS-1and ETS-2transcription factors.3. liver-enriched transcription factors C/EBP andHNF3play an important role in the transcriptional regulation of ApoF. |
