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The potential of the cruciform structure formation as an important factor influencing p53 sequence-specific binding to natural DNA targets
EB Jagelska, H Pivonkova, M Fojta, V Brazda
Language English Country United States
Document type Research Support, Non-U.S. Gov't
- MeSH
- Cell Line MeSH
- Enzyme-Linked Immunosorbent Assay MeSH
- Cyclin-Dependent Kinase Inhibitor p21 genetics MeSH
- Intracellular Signaling Peptides and Proteins genetics MeSH
- DNA, Single-Stranded chemistry metabolism MeSH
- Nucleic Acid Conformation MeSH
- Humans MeSH
- Tumor Suppressor Protein p53 genetics metabolism MeSH
- Inverted Repeat Sequences MeSH
- Plasmids chemistry metabolism MeSH
- Proto-Oncogene Proteins c-mdm2 genetics MeSH
- Gene Expression Regulation MeSH
- Electrophoretic Mobility Shift Assay MeSH
- Base Sequence MeSH
- DNA, Superhelical chemistry metabolism MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
p53 is one of the most important tumor suppressors which responds to DNA damage by binding to DNA and regulating the transcription of genes involved in cell cycle arrest, apoptosis, or senescence. As it was shown previously, p53 binding to DNA is strongly influenced by DNA topology. DNA supercoiling is fundamentally important for a wide range of biological processes including DNA transcription, replication, recombination, control of gene expression and genome organization. In this study, we investigated the cruciform structures formation of various inverted repeats in p53-responsive sequences from p21, RGC, mdm2 and GADD45 promoters under negative superhelical stress, and analyzed the effects of these DNA topology changes on p53-DNA binding. We demonstrated using three different methods (gel retardation analyses, ELISA and magnetic immunoprecipitation assay) that the p53 protein binds preferentially to negatively supercoiled plasmid DNAs with p53-responsive sequence presented as a cruciform structure. Not only the appearance of the cruciform structures within naked supercoiled DNA, but also the potential of the binding sites for adopting the non-B structures can contribute to a more favorable p53-DNA complex. Copyright 2009 Elsevier Inc. All rights reserved.
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- $a p53 is one of the most important tumor suppressors which responds to DNA damage by binding to DNA and regulating the transcription of genes involved in cell cycle arrest, apoptosis, or senescence. As it was shown previously, p53 binding to DNA is strongly influenced by DNA topology. DNA supercoiling is fundamentally important for a wide range of biological processes including DNA transcription, replication, recombination, control of gene expression and genome organization. In this study, we investigated the cruciform structures formation of various inverted repeats in p53-responsive sequences from p21, RGC, mdm2 and GADD45 promoters under negative superhelical stress, and analyzed the effects of these DNA topology changes on p53-DNA binding. We demonstrated using three different methods (gel retardation analyses, ELISA and magnetic immunoprecipitation assay) that the p53 protein binds preferentially to negatively supercoiled plasmid DNAs with p53-responsive sequence presented as a cruciform structure. Not only the appearance of the cruciform structures within naked supercoiled DNA, but also the potential of the binding sites for adopting the non-B structures can contribute to a more favorable p53-DNA complex. Copyright 2009 Elsevier Inc. All rights reserved.
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