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Preferential binding of hot spot mutant p53 proteins to supercoiled DNA in vitro and in cells

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Preferential binding of hot spot mutant p53 proteins to supercoiled DNA in vitro and in cells. / Brázdová, Marie; Navrátilová, Lucie; Tichý, Vlastimil; Němcová, Kateřina; Lexa, Matej; Hrstka, Roman; Pečinka, Petr; Adámik, Matej; Vojtesek, Borivoj; Paleček, Emil; Deppert, Wolfgang; Fojta, Miroslav.

in: PLOS ONE, Jahrgang 8, Nr. 3, 01.01.2013, S. e59567.

Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung

Harvard

Brázdová, M, Navrátilová, L, Tichý, V, Němcová, K, Lexa, M, Hrstka, R, Pečinka, P, Adámik, M, Vojtesek, B, Paleček, E, Deppert, W & Fojta, M 2013, 'Preferential binding of hot spot mutant p53 proteins to supercoiled DNA in vitro and in cells', PLOS ONE, Jg. 8, Nr. 3, S. e59567. https://doi.org/10.1371/journal.pone.0059567

APA

Brázdová, M., Navrátilová, L., Tichý, V., Němcová, K., Lexa, M., Hrstka, R., Pečinka, P., Adámik, M., Vojtesek, B., Paleček, E., Deppert, W., & Fojta, M. (2013). Preferential binding of hot spot mutant p53 proteins to supercoiled DNA in vitro and in cells. PLOS ONE, 8(3), e59567. https://doi.org/10.1371/journal.pone.0059567

Vancouver

Brázdová M, Navrátilová L, Tichý V, Němcová K, Lexa M, Hrstka R et al. Preferential binding of hot spot mutant p53 proteins to supercoiled DNA in vitro and in cells. PLOS ONE. 2013 Jan 1;8(3):e59567. https://doi.org/10.1371/journal.pone.0059567

Bibtex

@article{3bd0e5eb9ca1426cbe81355057696e46,
title = "Preferential binding of hot spot mutant p53 proteins to supercoiled DNA in vitro and in cells",
abstract = "Hot spot mutant p53 (mutp53) proteins exert oncogenic gain-of-function activities. Binding of mutp53 to DNA is assumed to be involved in mutp53-mediated repression or activation of several mutp53 target genes. To investigate the importance of DNA topology on mutp53-DNA recognition in vitro and in cells, we analyzed the interaction of seven hot spot mutp53 proteins with topologically different DNA substrates (supercoiled, linear and relaxed) containing and/or lacking mutp53 binding sites (mutp53BS) using a variety of electrophoresis and immunoprecipitation based techniques. All seven hot spot mutp53 proteins (R175H, G245S, R248W, R249S, R273C, R273H and R282W) were found to have retained the ability of wild-type p53 to preferentially bind circular DNA at native negative superhelix density, while linear or relaxed circular DNA was a poor substrate. The preference of mutp53 proteins for supercoiled DNA (supercoil-selective binding) was further substantiated by competition experiments with linear DNA or relaxed DNA in vitro and ex vivo. Using chromatin immunoprecipitation, the preferential binding of mutp53 to a sc mutp53BS was detected also in cells. Furthermore, we have shown by luciferase reporter assay that the DNA topology influences p53 regulation of BAX and MSP/MST1 promoters. Possible modes of mutp53 binding to topologically constrained DNA substrates and their biological consequences are discussed.",
keywords = "Binding Sites, Cell Line, Tumor, DNA, Superhelical, Gene Expression Regulation, Humans, Mutant Proteins, Mutation, Plasmids, Promoter Regions, Genetic, Protein Binding, Protein-Serine-Threonine Kinases, Substrate Specificity, Tumor Suppressor Protein p53, bcl-2-Associated X Protein",
author = "Marie Br{\'a}zdov{\'a} and Lucie Navr{\'a}tilov{\'a} and Vlastimil Tich{\'y} and Kate{\v r}ina N{\v e}mcov{\'a} and Matej Lexa and Roman Hrstka and Petr Pe{\v c}inka and Matej Ad{\'a}mik and Borivoj Vojtesek and Emil Pale{\v c}ek and Wolfgang Deppert and Miroslav Fojta",
year = "2013",
month = jan,
day = "1",
doi = "10.1371/journal.pone.0059567",
language = "English",
volume = "8",
pages = "e59567",
journal = "PLOS ONE",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "3",

}

RIS

TY - JOUR

T1 - Preferential binding of hot spot mutant p53 proteins to supercoiled DNA in vitro and in cells

AU - Brázdová, Marie

AU - Navrátilová, Lucie

AU - Tichý, Vlastimil

AU - Němcová, Kateřina

AU - Lexa, Matej

AU - Hrstka, Roman

AU - Pečinka, Petr

AU - Adámik, Matej

AU - Vojtesek, Borivoj

AU - Paleček, Emil

AU - Deppert, Wolfgang

AU - Fojta, Miroslav

PY - 2013/1/1

Y1 - 2013/1/1

N2 - Hot spot mutant p53 (mutp53) proteins exert oncogenic gain-of-function activities. Binding of mutp53 to DNA is assumed to be involved in mutp53-mediated repression or activation of several mutp53 target genes. To investigate the importance of DNA topology on mutp53-DNA recognition in vitro and in cells, we analyzed the interaction of seven hot spot mutp53 proteins with topologically different DNA substrates (supercoiled, linear and relaxed) containing and/or lacking mutp53 binding sites (mutp53BS) using a variety of electrophoresis and immunoprecipitation based techniques. All seven hot spot mutp53 proteins (R175H, G245S, R248W, R249S, R273C, R273H and R282W) were found to have retained the ability of wild-type p53 to preferentially bind circular DNA at native negative superhelix density, while linear or relaxed circular DNA was a poor substrate. The preference of mutp53 proteins for supercoiled DNA (supercoil-selective binding) was further substantiated by competition experiments with linear DNA or relaxed DNA in vitro and ex vivo. Using chromatin immunoprecipitation, the preferential binding of mutp53 to a sc mutp53BS was detected also in cells. Furthermore, we have shown by luciferase reporter assay that the DNA topology influences p53 regulation of BAX and MSP/MST1 promoters. Possible modes of mutp53 binding to topologically constrained DNA substrates and their biological consequences are discussed.

AB - Hot spot mutant p53 (mutp53) proteins exert oncogenic gain-of-function activities. Binding of mutp53 to DNA is assumed to be involved in mutp53-mediated repression or activation of several mutp53 target genes. To investigate the importance of DNA topology on mutp53-DNA recognition in vitro and in cells, we analyzed the interaction of seven hot spot mutp53 proteins with topologically different DNA substrates (supercoiled, linear and relaxed) containing and/or lacking mutp53 binding sites (mutp53BS) using a variety of electrophoresis and immunoprecipitation based techniques. All seven hot spot mutp53 proteins (R175H, G245S, R248W, R249S, R273C, R273H and R282W) were found to have retained the ability of wild-type p53 to preferentially bind circular DNA at native negative superhelix density, while linear or relaxed circular DNA was a poor substrate. The preference of mutp53 proteins for supercoiled DNA (supercoil-selective binding) was further substantiated by competition experiments with linear DNA or relaxed DNA in vitro and ex vivo. Using chromatin immunoprecipitation, the preferential binding of mutp53 to a sc mutp53BS was detected also in cells. Furthermore, we have shown by luciferase reporter assay that the DNA topology influences p53 regulation of BAX and MSP/MST1 promoters. Possible modes of mutp53 binding to topologically constrained DNA substrates and their biological consequences are discussed.

KW - Binding Sites

KW - Cell Line, Tumor

KW - DNA, Superhelical

KW - Gene Expression Regulation

KW - Humans

KW - Mutant Proteins

KW - Mutation

KW - Plasmids

KW - Promoter Regions, Genetic

KW - Protein Binding

KW - Protein-Serine-Threonine Kinases

KW - Substrate Specificity

KW - Tumor Suppressor Protein p53

KW - bcl-2-Associated X Protein

U2 - 10.1371/journal.pone.0059567

DO - 10.1371/journal.pone.0059567

M3 - SCORING: Journal article

C2 - 23555710

VL - 8

SP - e59567

JO - PLOS ONE

JF - PLOS ONE

SN - 1932-6203

IS - 3

ER -