Involvement of upstream stimulatory factor in regulation of the mouse Prnd gene coding for Doppel protein.
Promoter of the Prnd gene coding for the Prion-like protein Doppel contains two critical cis-regulatory elements, NF-Y consensus motif and canonical E-box. Here, we studied a role of the upstream stimulatory factor (USF) in the E-box-mediated activation of Prnd transcription. Co-expression of USF-1 with the luciferase reporter gene driven by the -185/+27 Prnd promoter fragment resulted in several fold increase of the luciferase activity. Conversely, mutations within the E-box led to a significantly reduced Prnd promoter activation. USF-1 binding was supported by the gel shift assay, supershift with USF-1 antibody and UV cross-linking. The activation capacity of the related USF-2, c-Myc and HIF-2alpha proteins was lower compared to USF-1 suggesting that USF-1 is the major E-box-binding transcription factor regulating the Prnd promoter.
Sepelakova
Janka
J
Centre of Molecular Medicine, Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9, Bratislava 845 05, Slovakia.
Takacova
Martina
M
Pastorekova
Silvia
S
Kopacek
Juraj
J
D000818:Animals; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D024721:E-Box Elements / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D058851:GPI-Linked Proteins; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D051379:Mice; D009154:Mutation; D011328:Prions / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D051720:Upstream Stimulatory Factors / Q000235:genetics* / Q000378:metabolism
D004268:DNA-Binding Proteins; D058851:GPI-Linked Proteins; D011328:Prions; C122036:Prnd protein, mouse; D014157:Transcription Factors; D051720:Upstream Stimulatory Factors; C497183:Usf1 protein, mouse