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Direct observation of damage clustering in irradiated DNA with atomic force microscopy

Ionizing radiation produces clustered DNA damage that contains two or more lesions in 10–20 bp. It is believed that the complexity of clustered damage (i.e., the number of lesions per damage site) is related to the biological severity of ionizing radiation. However, only simple clustered damage cont...

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Autores principales: Xu, Xu, Nakano, Toshiaki, Tsuda, Masataka, Kanamoto, Ryota, Hirayama, Ryoichi, Uzawa, Akiko, Ide, Hiroshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7026632/
https://www.ncbi.nlm.nih.gov/pubmed/31840169
http://dx.doi.org/10.1093/nar/gkz1159
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author Xu, Xu
Nakano, Toshiaki
Tsuda, Masataka
Kanamoto, Ryota
Hirayama, Ryoichi
Uzawa, Akiko
Ide, Hiroshi
author_facet Xu, Xu
Nakano, Toshiaki
Tsuda, Masataka
Kanamoto, Ryota
Hirayama, Ryoichi
Uzawa, Akiko
Ide, Hiroshi
author_sort Xu, Xu
collection PubMed
description Ionizing radiation produces clustered DNA damage that contains two or more lesions in 10–20 bp. It is believed that the complexity of clustered damage (i.e., the number of lesions per damage site) is related to the biological severity of ionizing radiation. However, only simple clustered damage containing two vicinal lesions has been demonstrated experimentally. Here we developed a novel method to analyze the complexity of clustered DNA damage. Plasmid DNA was irradiated with densely and sparsely ionizing Fe-ion beams and X-rays, respectively. Then, the resulting DNA lesions were labeled with biotin/streptavidin and observed with atomic force microscopy. Fe-ion beams produced complex clustered damage containing 2–4 lesions. Furthermore, they generated two or three clustered damage sites in a single plasmid molecule that resulted from the hit of a single track of Fe-ion beams. Conversely, X-rays produced relatively simple clustered damage. The present results provide the first experimental evidence for complex cluster damage.
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spelling pubmed-70266322020-02-25 Direct observation of damage clustering in irradiated DNA with atomic force microscopy Xu, Xu Nakano, Toshiaki Tsuda, Masataka Kanamoto, Ryota Hirayama, Ryoichi Uzawa, Akiko Ide, Hiroshi Nucleic Acids Res Methods Online Ionizing radiation produces clustered DNA damage that contains two or more lesions in 10–20 bp. It is believed that the complexity of clustered damage (i.e., the number of lesions per damage site) is related to the biological severity of ionizing radiation. However, only simple clustered damage containing two vicinal lesions has been demonstrated experimentally. Here we developed a novel method to analyze the complexity of clustered DNA damage. Plasmid DNA was irradiated with densely and sparsely ionizing Fe-ion beams and X-rays, respectively. Then, the resulting DNA lesions were labeled with biotin/streptavidin and observed with atomic force microscopy. Fe-ion beams produced complex clustered damage containing 2–4 lesions. Furthermore, they generated two or three clustered damage sites in a single plasmid molecule that resulted from the hit of a single track of Fe-ion beams. Conversely, X-rays produced relatively simple clustered damage. The present results provide the first experimental evidence for complex cluster damage. Oxford University Press 2020-02-20 2019-12-16 /pmc/articles/PMC7026632/ /pubmed/31840169 http://dx.doi.org/10.1093/nar/gkz1159 Text en © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Methods Online
Xu, Xu
Nakano, Toshiaki
Tsuda, Masataka
Kanamoto, Ryota
Hirayama, Ryoichi
Uzawa, Akiko
Ide, Hiroshi
Direct observation of damage clustering in irradiated DNA with atomic force microscopy
title Direct observation of damage clustering in irradiated DNA with atomic force microscopy
title_full Direct observation of damage clustering in irradiated DNA with atomic force microscopy
title_fullStr Direct observation of damage clustering in irradiated DNA with atomic force microscopy
title_full_unstemmed Direct observation of damage clustering in irradiated DNA with atomic force microscopy
title_short Direct observation of damage clustering in irradiated DNA with atomic force microscopy
title_sort direct observation of damage clustering in irradiated dna with atomic force microscopy
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7026632/
https://www.ncbi.nlm.nih.gov/pubmed/31840169
http://dx.doi.org/10.1093/nar/gkz1159
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