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Nouvelles sondes oligonucléotidiques fluorescentes ou paramagnétiques : applications à l'étude structurale des lésions de l'ADN et à leur réparation sur support.

Mélanie Flaender 1
1 LAN - Laboratoire Lésions des Acides Nucléiques
SCIB - UMR E3 - Service de Chimie Inorganique et Biologique : DSM/INAC/SCIB
Abstract : DNA which is the support of genetic information is constantly subjected to damaging agents. These processes can lead to structural modifications of the DNA molecule and harmful biological consequences, such as mutagenesis or cancer. DNA damages can be repaired by enzymatic complexes which restore the original sequence of the biopolymer. In this study we were interested in structural aspects of DNA lesions and in their repair by the base excision repair (BER) and the reversion repair (RR) pathways. The present work consisted in developing a new tool in a biochip format aim at detecting these repairs activities by fluorescence measurement. Thus, hairpin-shaped damaged oligonucleotides were grafted on glass slides by a click chemistry approach. After optimization of the immobilization conditions, by click chemistry, this new biosensor was validated for the detection of repair activities using purified enzymes (glycosylases and AP-endonucleases) and cell extracts. Using a similar approach, we adapted this microarray to measure reversion repair and to screen DNA repair inhibitors. In a second part of this work, we applied the pulsed electron paramagnetic resonance (pulsed EPR) technique to study structural deformation induced by several DNA damages. To achieve this goal we developed a multiple site-specific labelling procedure of the DNA probes by nitroxide radicals. Then, pulsed EPR was used for the first time to detect an enzymatic DNA repair activity.
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https://tel.archives-ouvertes.fr/tel-00558563
Contributor : Mélanie Flaender <>
Submitted on : Saturday, January 22, 2011 - 11:11:19 AM
Last modification on : Tuesday, May 11, 2021 - 11:36:07 AM
Long-term archiving on: : Saturday, April 23, 2011 - 2:42:47 AM

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  • HAL Id : tel-00558563, version 1

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Mélanie Flaender. Nouvelles sondes oligonucléotidiques fluorescentes ou paramagnétiques : applications à l'étude structurale des lésions de l'ADN et à leur réparation sur support.. Biochimie [q-bio.BM]. Université Joseph-Fourier - Grenoble I, 2010. Français. ⟨tel-00558563⟩

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