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:
Czech title
p53 se specificky váže na triplexy v DNA in vitro a v buňkách
journal article
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)

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


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.

PLoS ONE, vol. 11, no. 12, ISSN 1932-6203
Public Library of Science
EID Scopus
   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 = ""
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