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UV radiation effects on a DNA repair enzyme: conversion of a [4Fe–4S]2+ cluster into a [2Fe–2S]2+

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Abstract

Organisms are often exposed to different types of ionizing radiation that, directly or not, will promote damage to DNA molecules and/or other cellular structures. Because of that, organisms developed a wide range of response mechanisms to deal with these threats. Endonuclease III is one of the enzymes responsible to detect and repair oxidized pyrimidine base lesions. However, the effect of radiation on the structure/function of these enzymes is not clear yet. Here, we demonstrate the effect of UV-C radiation on E. coli endonuclease III through several techniques, namely UV–visible, fluorescence and Mössbauer spectroscopies, as well as SDS-PAGE and electrophoretic mobility shift assay. We demonstrate that irradiation with a UV-C source has dramatic consequences on the absorption, fluorescence, structure and functionality of the protein, affecting its [4Fe–4S] cluster and its DNA-binding ability, which results in its inactivation. An UV-C radiation-induced conversion of the [4Fe–4S]2+ into a [2Fe–2S]2+ was observed for the first time and proven by Mössbauer and UV–visible analysis. This work also shows that the DNA-binding capability of endonuclease III is highly dependent of the nuclearity of the endogenous iron–sulfur cluster. Thus, from our point of view, in a cellular context, these results strengthen the argument that cellular sensitivity to radiation can also be due to loss of radiation-induced damage repair ability.

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Acknowledgments

This research was supported by Fundação para a Ciência e Tecnologia, Ministério da Educação e Ciência, grant PTDC/SAU-SAP/111482/2009 (P.T.), PTDC/QUI/64248/2006 (to A.S.P), grant SFRH/BPD/48430/2008, a Post-Doctoral grant to F.F. Requimte was funded in part by grant PEst-C/EQB/LA0006/2011 from FCT/MEC.

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Authors and Affiliations

  1. REQUIMTE/CQFB, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829‐516, Caparica, Portugal

    Filipe Folgosa, Inês Camacho, Daniela Penas, Márcia Guilherme, João Fróis, Pedro Tavares & Alice S. Pereira

  2. CEFITEC, Departamento de Física, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal

    Filipe Folgosa & Paulo A. Ribeiro

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  1. Filipe Folgosa
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  2. Inês Camacho
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  3. Daniela Penas
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  4. Márcia Guilherme
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  8. Alice S. Pereira
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Correspondence to Alice S. Pereira.

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Folgosa, F., Camacho, I., Penas, D. et al. UV radiation effects on a DNA repair enzyme: conversion of a [4Fe–4S]2+ cluster into a [2Fe–2S]2+ . Radiat Environ Biophys 54, 111–121 (2015). https://doi.org/10.1007/s00411-014-0569-y

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