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New Environment-Sensitive Multichannel DNA Fluorescent Label for Investigation of the Protein-DNA Interactions

Here, we report the study of a new multichannel DNA fluorescent base analogue 3-hydroxychromone (3HC) to evaluate its suitability as a fluorescent reporter probe of structural transitions during protein-DNA interactions and its comparison with the current commercially available 2-aminopurine (aPu),...

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Autores principales: Kuznetsova, Alexandra A., Kuznetsov, Nikita A., Vorobjev, Yuri N., Barthes, Nicolas P. F., Michel, Benoît Y., Burger, Alain, Fedorova, Olga S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4055743/
https://www.ncbi.nlm.nih.gov/pubmed/24925085
http://dx.doi.org/10.1371/journal.pone.0100007
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author Kuznetsova, Alexandra A.
Kuznetsov, Nikita A.
Vorobjev, Yuri N.
Barthes, Nicolas P. F.
Michel, Benoît Y.
Burger, Alain
Fedorova, Olga S.
author_facet Kuznetsova, Alexandra A.
Kuznetsov, Nikita A.
Vorobjev, Yuri N.
Barthes, Nicolas P. F.
Michel, Benoît Y.
Burger, Alain
Fedorova, Olga S.
author_sort Kuznetsova, Alexandra A.
collection PubMed
description Here, we report the study of a new multichannel DNA fluorescent base analogue 3-hydroxychromone (3HC) to evaluate its suitability as a fluorescent reporter probe of structural transitions during protein-DNA interactions and its comparison with the current commercially available 2-aminopurine (aPu), pyrrolocytosine (C(py)) and 1,3-diaza-2-oxophenoxazine (tC(O)). For this purpose, fluorescent base analogues were incorporated into DNA helix on the opposite or on the 5′-side of the damaged nucleoside 5,6-dihydrouridine (DHU), which is specifically recognized and removed by Endonuclease VIII. These fluorophores demonstrated different sensitivities to the DNA helix conformational changes. The highest sensitivity and the most detailed information about the conformational changes of DNA induced by protein binding and processing were obtained using the 3HC probe. The application of this new artificial fluorescent DNA base is a very useful tool for the studies of complex mechanisms of protein-DNA interactions. Using 3HC biosensor, the kinetic mechanism of Endonuclease VIII action was specified.
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spelling pubmed-40557432014-06-18 New Environment-Sensitive Multichannel DNA Fluorescent Label for Investigation of the Protein-DNA Interactions Kuznetsova, Alexandra A. Kuznetsov, Nikita A. Vorobjev, Yuri N. Barthes, Nicolas P. F. Michel, Benoît Y. Burger, Alain Fedorova, Olga S. PLoS One Research Article Here, we report the study of a new multichannel DNA fluorescent base analogue 3-hydroxychromone (3HC) to evaluate its suitability as a fluorescent reporter probe of structural transitions during protein-DNA interactions and its comparison with the current commercially available 2-aminopurine (aPu), pyrrolocytosine (C(py)) and 1,3-diaza-2-oxophenoxazine (tC(O)). For this purpose, fluorescent base analogues were incorporated into DNA helix on the opposite or on the 5′-side of the damaged nucleoside 5,6-dihydrouridine (DHU), which is specifically recognized and removed by Endonuclease VIII. These fluorophores demonstrated different sensitivities to the DNA helix conformational changes. The highest sensitivity and the most detailed information about the conformational changes of DNA induced by protein binding and processing were obtained using the 3HC probe. The application of this new artificial fluorescent DNA base is a very useful tool for the studies of complex mechanisms of protein-DNA interactions. Using 3HC biosensor, the kinetic mechanism of Endonuclease VIII action was specified. Public Library of Science 2014-06-12 /pmc/articles/PMC4055743/ /pubmed/24925085 http://dx.doi.org/10.1371/journal.pone.0100007 Text en © 2014 Kuznetsova et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Kuznetsova, Alexandra A.
Kuznetsov, Nikita A.
Vorobjev, Yuri N.
Barthes, Nicolas P. F.
Michel, Benoît Y.
Burger, Alain
Fedorova, Olga S.
New Environment-Sensitive Multichannel DNA Fluorescent Label for Investigation of the Protein-DNA Interactions
title New Environment-Sensitive Multichannel DNA Fluorescent Label for Investigation of the Protein-DNA Interactions
title_full New Environment-Sensitive Multichannel DNA Fluorescent Label for Investigation of the Protein-DNA Interactions
title_fullStr New Environment-Sensitive Multichannel DNA Fluorescent Label for Investigation of the Protein-DNA Interactions
title_full_unstemmed New Environment-Sensitive Multichannel DNA Fluorescent Label for Investigation of the Protein-DNA Interactions
title_short New Environment-Sensitive Multichannel DNA Fluorescent Label for Investigation of the Protein-DNA Interactions
title_sort new environment-sensitive multichannel dna fluorescent label for investigation of the protein-dna interactions
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4055743/
https://www.ncbi.nlm.nih.gov/pubmed/24925085
http://dx.doi.org/10.1371/journal.pone.0100007
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