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Interaction entre yOgg1, une ADN glycosylase de la voie BER, et l’ADN polymérase réplicative Polε chez Saccharomyces cerevisiae

Abstract : Oxidative DNA damages are involved in pathological processes such as cancer, neurodegenerative diseases and aging. Part of these damages results from the action of reactive oxygen species (ROS), which are produced by cellular metabolism or (physical or chemical) exogenous agents. They lead to different types of DNA lesions including DNA base oxidation (8-oxoguanine, 8-oxoG) and abasic site formation (AP, apuric/apyrimidic). If not removed, these lesions lead to mutagenesis or cell death. Most of base lesions are dealt specifically by the base excision repair (BER) pathway. BER is initiated by a DNA glycosylase, such as 8-oxoG-DNA glycosylase (Ogg1) which is responsible for the removal of 8-oxoG. In previous unpublished work, a yeast two-hybrid study revealed the existence in S. cerevisiae of an interaction between yOgg1 and the catalytic subunit of the replicative DNA polymerase Polε (yPol2), also involved in the BER pathway in eukaryotes. Our work shows that yOgg1 and yPol2 physically and specifically interact with each other. Truncation and site-directed mutagenesis studies allowed us to identify the 3 ' → 5' exonuclease activity domain of yPol2 as part of the minimal form of yPol2 still able to interact with yOgg1. The active site of yOgg1 and/or its immediate vicinity may contain part of its interaction domain with yPol2. Besides, we observe a clear correlation between yOgg1 catalytic activity and its ability to interact with yPol2 in vivo. Similarly, the 3'→5' exonuclease activity of yPol2 could be useful to its interaction with yOgg1. From a functional point of view, yPol2 stimulates in vitro the AP lyase activity of yOgg1 and the coupling of both DNA glycosylase and AP lyase enzyme activity. The interaction yOgg1/yPol2 could allow a better coordination of damaged nucleoside excision and DNA re-synthesis steps in BER.
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Submitted on : Thursday, April 23, 2015 - 3:42:05 PM
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Kadija Essalhi. Interaction entre yOgg1, une ADN glycosylase de la voie BER, et l’ADN polymérase réplicative Polε chez Saccharomyces cerevisiae. Sciences agricoles. Université d'Orléans, 2013. Français. ⟨NNT : 2013ORLE2077⟩. ⟨tel-01145247⟩



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