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UVA Irradiation of BrU-Substituted DNA in the Presence of Hoechst 33258

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Molecular Recognition of DNA Double Helix

Part of the book series: Springer Theses ((Springer Theses))

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Abstract

Given that our knowledge of DNA repair is limited because of the complexity of the DNA system, a technique called UVA micro-irradiation has been developed that can be used to visualize the recruitment of DNA repair proteins at double-strand break (DSB) sites. Interestingly, Hoechst 33258 was used under micro-irradiation to sensitize 5-bromouracil (BrU)-labeled DNA, causing efficient DSBs. However, the molecular basis of DSB formation under UVA micro-irradiation remains unknown. Herein, we investigated the mechanism of DSB formation under UVA micro-irradiation conditions. Our results suggest that the generation of a uracil-5-yl radical through electron transfer from Hoechst 33258 to BrU caused DNA cleavage preferentially at self-complementary 5′-AABrUBrU-3′ sequences to induce DSB. We also investigated the DNA cleavage in the context of the nucleosome to gain a better understanding of UVA micro-irradiation in a cell-like model. We found that DNA cleavage occurred in both core and linker DNA regions although its efficiency reduced in core DNA.

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  1. Institute Curie, Orsay, France

    Dr. Abhijit Saha

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  1. Dr. Abhijit Saha
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Correspondence to Abhijit Saha .

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Saha, A. (2018). UVA Irradiation of BrU-Substituted DNA in the Presence of Hoechst 33258. In: Molecular Recognition of DNA Double Helix. Springer Theses. Springer, Singapore. https://doi.org/10.1007/978-981-10-8746-2_6

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