Cargando…

TWJ-Screen: an isothermal screening assay to assess ligand/DNA junction interactions in vitro

The quest for chemicals able to operate at selected genomic loci in a spatiotemporally controlled manner is desirable to create manageable DNA damages. Mounting evidence now shows that alternative DNA structures, including G-quadruplexes and branched DNA (or DNA junctions), might hamper proper progr...

Descripción completa

Detalles Bibliográficos
Autores principales: Guyon, Ludivine, Pirrotta, Marc, Duskova, Katerina, Granzhan, Anton, Teulade-Fichou, Marie-Paule, Monchaud, David
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5815093/
https://www.ncbi.nlm.nih.gov/pubmed/29149299
http://dx.doi.org/10.1093/nar/gkx1118
_version_ 1783300447391449088
author Guyon, Ludivine
Pirrotta, Marc
Duskova, Katerina
Granzhan, Anton
Teulade-Fichou, Marie-Paule
Monchaud, David
author_facet Guyon, Ludivine
Pirrotta, Marc
Duskova, Katerina
Granzhan, Anton
Teulade-Fichou, Marie-Paule
Monchaud, David
author_sort Guyon, Ludivine
collection PubMed
description The quest for chemicals able to operate at selected genomic loci in a spatiotemporally controlled manner is desirable to create manageable DNA damages. Mounting evidence now shows that alternative DNA structures, including G-quadruplexes and branched DNA (or DNA junctions), might hamper proper progression of replication fork, thus triggering DNA damages and genomic instability. Therefore, small molecules that stabilize these DNA structures are currently scrutinized as a promising way to create genomic defects that cannot be dealt with properly by cancer cells. While much emphasis has been recently given to G-quadruplexes and related ligands, we report herein on three-way DNA junctions (TWJ) and related ligands. We first highlight the biological implications of TWJ and their strategic relevance as triggers for replicative stress. Then, we describe a new in vitro high-throughput screening assay, TWJ-Screen, which allows for identifying TWJ ligands with both high affinity and selectivity for TWJ over other DNA structures (duplexes and quadruplexes), in a convenient and unbiased manner as demonstrated by the screening of a library of 25 compounds from different chemical families. TWJ-Screen thus represents a reliable mean to uncover molecular tools able to foster replicative stress through an innovative approach, thus providing new strategic opportunities to combat cancers.
format Online
Article
Text
id pubmed-5815093
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher Oxford University Press
record_format MEDLINE/PubMed
spelling pubmed-58150932018-02-23 TWJ-Screen: an isothermal screening assay to assess ligand/DNA junction interactions in vitro Guyon, Ludivine Pirrotta, Marc Duskova, Katerina Granzhan, Anton Teulade-Fichou, Marie-Paule Monchaud, David Nucleic Acids Res Methods Online The quest for chemicals able to operate at selected genomic loci in a spatiotemporally controlled manner is desirable to create manageable DNA damages. Mounting evidence now shows that alternative DNA structures, including G-quadruplexes and branched DNA (or DNA junctions), might hamper proper progression of replication fork, thus triggering DNA damages and genomic instability. Therefore, small molecules that stabilize these DNA structures are currently scrutinized as a promising way to create genomic defects that cannot be dealt with properly by cancer cells. While much emphasis has been recently given to G-quadruplexes and related ligands, we report herein on three-way DNA junctions (TWJ) and related ligands. We first highlight the biological implications of TWJ and their strategic relevance as triggers for replicative stress. Then, we describe a new in vitro high-throughput screening assay, TWJ-Screen, which allows for identifying TWJ ligands with both high affinity and selectivity for TWJ over other DNA structures (duplexes and quadruplexes), in a convenient and unbiased manner as demonstrated by the screening of a library of 25 compounds from different chemical families. TWJ-Screen thus represents a reliable mean to uncover molecular tools able to foster replicative stress through an innovative approach, thus providing new strategic opportunities to combat cancers. Oxford University Press 2018-02-16 2017-11-15 /pmc/articles/PMC5815093/ /pubmed/29149299 http://dx.doi.org/10.1093/nar/gkx1118 Text en © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Methods Online
Guyon, Ludivine
Pirrotta, Marc
Duskova, Katerina
Granzhan, Anton
Teulade-Fichou, Marie-Paule
Monchaud, David
TWJ-Screen: an isothermal screening assay to assess ligand/DNA junction interactions in vitro
title TWJ-Screen: an isothermal screening assay to assess ligand/DNA junction interactions in vitro
title_full TWJ-Screen: an isothermal screening assay to assess ligand/DNA junction interactions in vitro
title_fullStr TWJ-Screen: an isothermal screening assay to assess ligand/DNA junction interactions in vitro
title_full_unstemmed TWJ-Screen: an isothermal screening assay to assess ligand/DNA junction interactions in vitro
title_short TWJ-Screen: an isothermal screening assay to assess ligand/DNA junction interactions in vitro
title_sort twj-screen: an isothermal screening assay to assess ligand/dna junction interactions in vitro
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5815093/
https://www.ncbi.nlm.nih.gov/pubmed/29149299
http://dx.doi.org/10.1093/nar/gkx1118
work_keys_str_mv AT guyonludivine twjscreenanisothermalscreeningassaytoassessliganddnajunctioninteractionsinvitro
AT pirrottamarc twjscreenanisothermalscreeningassaytoassessliganddnajunctioninteractionsinvitro
AT duskovakaterina twjscreenanisothermalscreeningassaytoassessliganddnajunctioninteractionsinvitro
AT granzhananton twjscreenanisothermalscreeningassaytoassessliganddnajunctioninteractionsinvitro
AT teuladefichoumariepaule twjscreenanisothermalscreeningassaytoassessliganddnajunctioninteractionsinvitro
AT monchauddavid twjscreenanisothermalscreeningassaytoassessliganddnajunctioninteractionsinvitro