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p53 Specifically Binds Triplex DNA In Vitro and in Cells
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SYSNO ASEP 0471941 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title p53 Specifically Binds Triplex DNA In Vitro and in Cells Author(s) Brázdová, Marie (BFU-R) RID, ORCID
Tichý, Vlastimil (BFU-R) RID
Helma, Robert (BFU-R) ORCID
Bažantová, Pavla (BFU-R)
Polášková, Alena (BFU-R)
Krejčí, A. (CZ)
Petr, Marek (BFU-R) ORCID
Navrátilová, Lucie (BFU-R)
Tichá, Olga (BFU-R)
Nejedlý, Karel (BFU-R) RID
Bennink, M.L. (CZ)
Subramaniam, V. (DE)
Babkova, Z. (CZ)
Martínek, T. (CZ)
Lexa, M. (CZ)
Adámik, Matěj (BFU-R) ORCIDNumber of authors 16 Article number e0167439 Source Title PLoS ONE. - : Public Library of Science - ISSN 1932-6203
Roč. 11, č. 12 (2016)Number of pages 25 s. Publication form Online - E Language eng - English Country US - United States Keywords c-terminal domain ; suppressor protein p53 ; supercoiled dna Subject RIV BO - Biophysics R&D Projects GA13-36108S GA ČR - Czech Science Foundation (CSF) GP204/06/P369 GA ČR - Czech Science Foundation (CSF) GA15-02891S GA ČR - Czech Science Foundation (CSF) Institutional support BFU-R - RVO:68081707 UT WOS 000389482700156 DOI 10.1371/journal.pone.0167439 Annotation Triplex DNA is implicated in a wide range of biological activities, including regulation of gene expression and genomic instability leading to cancer. The tumor suppressor p53 is a central regulator of cell fate in response to different type of insults. Sequence and structure specific modes of DNA recognition are core attributes of the p53 protein. The focus of this work is the structure-specific binding of p53 to DNA containing triplex-forming sequences in vitro and in cells and the effect on p53-driven transcription. This is the first DNA binding study of full-length p53 and its deletion variants to both intermolecular and intramolecular T.A.T triplexes. We demonstrate that the interaction of p53 with intermolecular T.A.T triplex is comparable to the recognition of CTG-hairpin non-B DNA structure. Using deletion mutants we determined the C-terminal DNA binding domain of p53 to be crucial for triplex recognition. Furthermore, strong p53 recognition of intramolecular T.A.T triplexes (H-DNA), stabilized by negative superhelicity in plasmid DNA, was detected by competition and immunoprecipitation experiments, and visualized by AFM. Moreover, chromatin immunoprecipitation revealed p53 binding T.A.T forming sequence in vivo. Enhanced reporter transactivation by p53 on insertion of triplex forming sequence into plasmid with p53 consensus sequence was observed by luciferase reporter assays. In-silico scan of human regulatory regions for the simultaneous presence of both consensus sequence and T.A.T motifs identified a set of candidate p53 target genes and p53-dependent activation of several of them (ABCG5, ENOX1, INSR, MCC, NFAT5) was confirmed by RT-qPCR. Our results show that T.A.T triplex comprises a new class of p53 binding sites targeted by p53 in a DNA structure-dependent mode in vitro and in cells. The contribution of p53 DNA structure-dependent binding to the regulation of transcription is discussed. Workplace Institute of Biophysics Contact Jana Poláková, polakova@ibp.cz, Tel.: 541 517 244 Year of Publishing 2017
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