yy1-binding sites provide central switch functions in the parp-1 gene expression networkyy1-binding网站提供中央开关功能的parp-1基因表达网络.pdf

YY1-Binding Sites Provide Central Switch Functions in the PARP-1 Gene Expression Network 1,2 1,3 ´4 1,5 ´ 1,4 Martina Doetsch , Angela Gluch , Goran Poznanovic , Juergen Bode , Melita Vidakovic * 1 Helmholtz Centre for Infection Research/Epigenetic Regulation, Braunschweig, Germany, 2 Department of Biochemistry and Molecular Cell Biology, Max F. Perutz Laboratories, University of Vienna, Vienna, Austria, 3 BIOBASE GmbH, Wolfenbuettel, Germany, 4 Department of Molecular Biology, Institute for Biological Research, University of Belgrade, Belgrade, Serbia, 5 Hannover Medical School (MHH), Experimental Hematology, Hannover, Germany Abstract Evidence is presented for the involvement of the interplay between transcription factor Yin Yang 1 (YY1) and poly(ADP- ribose) polymerase-1 (PARP-1) in the regulation of mouse PARP-1 gene (muPARP-1) promoter activity. We identified potential YY1 binding motifs (BM) at seven positions in the muPARP-1 core-promoter ( 2574/+200). Binding of YY1 was observed by the electrophoretic supershift assay using anti-YY1 antibody and linearized or supercoiled forms of plasmids bearing the core promoter, as well as with 30 bp oligonucleotide probes containing the individual YY1 binding motifs and four muPARP-1 promoter fragments. We detected YY1 binding to BM1 (2587/ 2558), BM4 (2348/2319) and a very prominent association with BM7 (+86/+ 115). Inspection of BM7 reveals overlap of the muPARP-1 translation start site with the Kozak sequence and YY1 and PARP-1 recognition sites. Site-directed mutagenesis of the YY1 and PARP-1 core motifs e