The Mechanism Of Fibroblast Growth Factor19 Transcriptional Regulation

ObjectiveThe expression level of FGF19 have been evaluated and the abnormity of FGF19 in a series of diseases,including type 2 diabetes and non alcoholic fatty liver disease,may play an important role in the pathophysiology progresss.In order to investigate the transcriptional regulation mechanism about FGF19,we constructed a series of plasmids containing the promoter fragments of FGF19 gene.MethodsIn order to get a full length of 3323 bp 5'-flanking fragment of FGF19,we projected specific primer to amplify the DNA sequence,and then the product was cloned into p GL3-Basic Vector.In view of the constructed promoter fragment of 3323 bp,several plasmids were established with mutation and deletion of 5' terminal.Subsequently,Transient transfection experiments were done to localize the core fragments of FGF19 promoter.The online software gene regulation,with the function of predicting transcription factor binding sites,identify latent transcriptional factors.Finally,we use EMSA to verify the binding of DNA fragments and latent transcription factors in vitro.Results1,we successfully cloned promoter plasmids of FGF19 gene verified by restriction endonuclease digestion and sequence analysis.2,In view of the transient transfection results of dual luciferase reporter plasmids,the key transcriptional regulation fragment was localized between-2351 to-2316 of FGF19 promoter.3,The analysis of FGF19 promoter fragments by gene regulation software identify the latent transcription factors.4,The position of-2236 and-2252 is Single-nucleotide polymorphism(SNP),and its mutation can regulate the activity of FGF19 promoter.EMSA verify the binding of DNA fragments with potent transcription factors in vitro.ConclusionThe fragment of FGF19 promoter from-2351 to-2316 is the core transcription regulation region and contain the pivotal binding sites of transcription factors,which may act as positive response elements in the promoter of FGF19.The binding of GR with fragemnts including-2236 can regulate the transcription of FGF19.