Publication detail

p53 Specifically Binds Triplex DNA In Vitro and in Cells

BRÁZDOVÁ, M. TICHÝ, V. HELMA, R. BAŽANTOVÁ, P. POLÁŠKOVÁ, A. KREJČÍ, A. PETR, M. NAVRÁTILOVÁ, L. TICHÁ, O. NEJEDLÝ, K. BENNINK, M. SUBRAMANIAM, V. BÁBKOVÁ, Z. MARTÍNEK, T. LEXA, M. ADÁMIK, M.

Original Title

p53 Specifically Binds Triplex DNA In Vitro and in Cells

Type

journal article in Web of Science

Language

English

Original 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.

Keywords

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

Authors

BRÁZDOVÁ, M.; TICHÝ, V.; HELMA, R.; BAŽANTOVÁ, P.; POLÁŠKOVÁ, A.; KREJČÍ, A.; PETR, M.; NAVRÁTILOVÁ, L.; TICHÁ, O.; NEJEDLÝ, K.; BENNINK, M.; SUBRAMANIAM, V.; BÁBKOVÁ, Z.; MARTÍNEK, T.; LEXA, M.; ADÁMIK, M.

Released

1. 12. 2016

ISBN

1932-6203

Periodical

PLOS ONE

Year of study

11

Number

12

State

United States of America

Pages from

1

Pages to

25

Pages count

25

URL

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0167439

BibTex

@article{BUT133510,
  author="Marie {Brázdová} and Vlastimil {Tichý} and Robert {Helma} and Pavla {Bažantová} and Alena {Polášková} and Aneta {Krejčí} and Marek {Petr} and Lucie {Navrátilová} and Olga {Tichá} and Karel {Nejedlý} and Martin {Bennink} and Vinod {Subramaniam} and Zuzana {Bábková} and Tomáš {Martínek} and Matej {Lexa} and Matej {Adámik}",
  title="p53 Specifically Binds Triplex DNA In Vitro and in Cells",
  journal="PLOS ONE",
  year="2016",
  volume="11",
  number="12",
  pages="1--25",
  doi="10.1371/journal.pone.0167439",
  issn="1932-6203",
  url="http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0167439"
}