The DNA damage response (DDR) has a critical role in the maintenance of genomic integrity during chromosome replication. However, responses to replication stress evoked by tight DNA–protein complexes have not been fully elucidated. Here, we used bacterial LacI protein binding to lacO arrays to make site-specific replication fork barriers on the human chromosome. These barriers induced the accumulation of single-stranded DNA (ssDNA) and various DDR proteins at the lacO site. SLX4–XPF functioned as an upstream factor for the accumulation of DDR proteins, and consequently, ATR and FANCD2 were interdependently recruited. Moreover, LacI binding in S phase caused underreplication and abnormal mitotic segregation of the lacO arrays. Finally, we show that the SLX4–ATR axis represses the anaphase abnormality induced by LacI binding. Our results outline a long-term process by which human cells manage nucleoprotein obstacles ahead of the replication fork to prevent chromosomal instability.
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4 January 2021
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December 21 2020
SLX4–XPF mediates DNA damage responses to replication stress induced by DNA–protein interactions
Riko Ishimoto
,
Riko Ishimoto
*
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
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Yota Tsuzuki
,
Yota Tsuzuki
*
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
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Tomoki Matsumura
,
Tomoki Matsumura
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
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Seiichiro Kurashige
,
Seiichiro Kurashige
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
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Kouki Enokitani
,
Kouki Enokitani
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
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Koki Narimatsu
,
Koki Narimatsu
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
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Mitsunori Higa
,
Mitsunori Higa
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
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Nozomi Sugimoto
,
Nozomi Sugimoto
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
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Kazumasa Yoshida
,
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
Correspondence to Kazumasa Yoshida: kyoshida@phar.kyushu-u.ac.jp
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Masatoshi Fujita
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
Masatoshi Fujita: mfujita@phar.kyushu-u.ac.jp
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Riko Ishimoto
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
Yota Tsuzuki
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
Tomoki Matsumura
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
Seiichiro Kurashige
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
Kouki Enokitani
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
Koki Narimatsu
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
Mitsunori Higa
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
Nozomi Sugimoto
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
Kazumasa Yoshida
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
Masatoshi Fujita
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
Correspondence to Kazumasa Yoshida: kyoshida@phar.kyushu-u.ac.jp
Masatoshi Fujita: mfujita@phar.kyushu-u.ac.jp
*
R. Ishimoto and Y. Tsuzuki contributed equally to this paper.
Received:
March 23 2020
Revision Received:
October 05 2020
Accepted:
November 13 2020
Online Issn: 1540-8140
Print Issn: 0021-9525
Funding:
Fukuoka Foundation for Sound Health
(NO AWARD)
Japan Society for the Promotion of Science London
(15K18478)
Mochida Memorial Foundation for Medical and Pharmaceutical Research
(NO AWARD)
© 2020 Ishimoto et al.
2020
This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).
J Cell Biol (2021) 220 (1): e202003148.
Article history
Received:
March 23 2020
Revision Received:
October 05 2020
Accepted:
November 13 2020
Citation
Riko Ishimoto, Yota Tsuzuki, Tomoki Matsumura, Seiichiro Kurashige, Kouki Enokitani, Koki Narimatsu, Mitsunori Higa, Nozomi Sugimoto, Kazumasa Yoshida, Masatoshi Fujita; SLX4–XPF mediates DNA damage responses to replication stress induced by DNA–protein interactions. J Cell Biol 4 January 2021; 220 (1): e202003148. doi: https://doi.org/10.1083/jcb.202003148
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