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Updated: May 19, 2026
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PAR-CliP - A Method to Identify Transcriptome-wide the Binding Sites of RNA Binding Proteins
Published on: July 2, 2010
YY1-binding sites provide central switch functions in the PARP-1 gene expression network
Martina Doetsch1, Angela Gluch, Goran Poznanović
1Helmholtz Centre for Infection Research/Epigenetic Regulation, Braunschweig, Germany.
Plos One
|September 1, 2012
Summary
The interplay between transcription factor Yin Yang 1 (YY1) and poly(ADP-ribose) polymerase-1 (PARP-1) regulates mouse PARP-1 gene expression. YY1 binding to specific promoter sites represses muPARP-1, while YY1 and PARP-1 show synergistic action.
Area of Science:
- Molecular Biology
- Gene Regulation
- Biochemistry
Background:
- Poly(ADP-ribose) polymerase-1 (PARP-1) plays a crucial role in DNA repair and gene expression.
- Transcription factors, such as Yin Yang 1 (YY1), are key regulators of gene promoter activity.
Purpose of the Study:
- To investigate the involvement of YY1 in the regulation of mouse PARP-1 (muPARP-1) gene promoter activity.
- To elucidate the interplay between YY1 and PARP-1 in controlling muPARP-1 expression.
Main Methods:
- Identification of YY1 binding motifs (BM) in the muPARP-1 core-promoter using bioinformatics.
- Electrophoretic supershift assays and oligonucleotide probes to confirm YY1 binding to specific motifs.
- Site-directed mutagenesis to assess the functional impact of YY1 and PARP-1 binding sites.
- Reporter gene transfection assays to evaluate promoter activity.
- Experiments in PARP-1 knockout cells with PARP-1 overexpression.
Main Results:
- Seven potential YY1 binding motifs were identified in the muPARP-1 promoter region.
- YY1 was confirmed to bind to BM1, BM4, and prominently to BM7, which overlaps with the translation start site and Kozak sequence.
- Mutagenesis of YY1/PARP-1 motifs abolished protein binding and demonstrated YY1's role in mediating PARP-1 binding.
- YY1 binding to BM1 and BM4 repressed muPARP-1 promoter activity, an effect reversed by mutation.
- Synergistic action between YY1 and PARP-1 was observed, suggesting a coordinated regulatory mechanism.
Conclusions:
- YY1 directly interacts with the muPARP-1 promoter, repressing its activity through specific binding sites.
- YY1 mediates PARP-1 binding near the translation start site, expanding the PARP-1 autoregulatory loop.
- The combined actions of YY1 and PARP-1 contribute to the strict limitation of muPARP-1 expression and cellular homeostasis.
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