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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.
Originální název
p53 Specifically Binds Triplex DNA In Vitro and in Cells
Typ
článek v časopise ve Web of Science, Jimp
Jazyk
angličtina
Originální abstrakt
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.
Klíčová slova
p53; DNA structure; DNA triplex; protein-DNA interaction; transcription target gene; transcription regulation
Autoři
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.
Vydáno
1. 12. 2016
ISSN
1932-6203
Periodikum
PLOS ONE
Ročník
11
Číslo
12
Stát
Spojené státy americké
Strany od
1
Strany do
25
Strany počet
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" }