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Surface plasmon resonance imaging of the conversion of clustered DNA lesions into double strand breaks by Fpg protein

1 Univ. Grenoble Alpes, UMR 5250, F-38000 Grenoble, France CNRS, DCM UMR 5250, F-38000 Grenoble, France;
2 Univ. Grenoble Alpes, INAC, SPrAM, F-38000 Grenoble, France CEA, INAC, SPrAM, F-38000 Grenoble, France CNRS, SPrAM, F-38000 Grenoble, France;
3 Current address: DKMS German Bone Marrow Donor Center, 72072 Tübingen, Germany

Topical Section: Biomolecular Materials: DNA, RNA, proteins, peptides, lipids

Ionizing radiations induce clustered damage sites known to be severely challenging for the cell's repair machinery. In the present study, we have grafted on a biochip four synthetic oligonucleotide duplexes containing either a single 8-oxoguanine, a single 2'-deoxyribose abasic site or two different clustered damages containing these two lesions on opposite strands. Using a SPR imaging-based DNA array, we qualitatively observed the hierarchy of lesion processing by the base excision repair (BER) enzyme Fpg (Formamidopyrimidine (fapy)-DNA glycosylase). We could notably show that Fpg can convert a cluster of lesions into a potentially lethal double strand break (DSB) and demonstrate that SPR imaging is suitable to investigate the incision steps occurring during the repair process.
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