Publication Details

p53 Specifically Binds Triplex DNA In Vitro and in Cells

BRÁZDOVÁ Marie, TICHÝ Vlastimil, HELMA Robert, BAŽANTOVÁ Pavla, POLÁŠKOVÁ Alena, KREJČÍ Aneta, PETR Marek, NAVRÁTILOVÁ Lucie, TICHÁ Olga, NEJEDLÝ Karel, BENNINK Martin L., SUBRAMANIAM Vinod, BÁBKOVÁ Zuzana, MARTÍNEK Tomáš, LEXA Matej and ADÁMIK Matej. p53 Specifically Binds Triplex DNA In Vitro and in Cells. PLoS ONE, vol. 11, no. 12, 2016, pp. 1-25. ISSN 1932-6203. Available from: http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0167439
Czech title
p53 se specificky váže na triplexy v DNA in vitro a v buňkách
Type
journal article
Language
english
Authors
Brázdová Marie, Mgr., Ph.D. (IBP)
Tichý Vlastimil (IBP)
Helma Robert, Bc. (IBP)
Bažantová Pavla (IBP)
Polášková Alena (IBP)
Krejčí Aneta (VFU)
Petr Marek (CAS CR)
Navrátilová Lucie, Mgr. (IBP)
Tichá Olga (IBP)
Nejedlý Karel (IBP)
Bennink Martin L. (UTWENTE)
Subramaniam Vinod (UTWENTE)
Bábková Zuzana (VFU)
Martínek Tomáš, doc. Ing., Ph.D. (DCSY FIT BUT)
Lexa Matej, Ing., Ph.D. (FI MUNI)
Adámik Matej (IBP)
URL
Keywords

p53; DNA structure; DNA triplex; protein-DNA interaction; transcription target gene; transcription regulation

Abstract

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.

Published
2016
Pages
1-25
Journal
PLoS ONE, vol. 11, no. 12, ISSN 1932-6203
Publisher
Public Library of Science
DOI
10.1371/journal.pone.0167439
UT WoS
000389482700156
EID Scopus
2-s2.0-85000868554
BibTeX 
@ARTICLE{FITPUB11343,
   author = "Marie Br\'{a}zdov\'{a} and Vlastimil Tich\'{y} and Robert Helma and Pavla Ba\v{z}antov\'{a} and Alena Pol\'{a}\v{s}kov\'{a} and Aneta Krej\v{c}\'{i} and Marek Petr and Lucie Navr\'{a}tilov\'{a} and Olga Tich\'{a} and Karel Nejedl\'{y} and L. Martin Bennink and Vinod Subramaniam and Zuzana B\'{a}bkov\'{a} and Tom\'{a}\v{s} Mart\'{i}nek and Matej Lexa and Matej Ad\'{a}mik",
   title = "p53 Specifically Binds Triplex DNA In Vitro and in Cells",
   pages = "1--25",
   journal = "PLoS ONE",
   volume = 11,
   number = 12,
   year = 2016,
   ISSN = "1932-6203",
   doi = "10.1371/journal.pone.0167439",
   language = "english",
   url = "https://www.fit.vut.cz/research/publication/11343"
}
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