CDC6 interaction with ATR regulates activation of a replication checkpoint in higher eukaryotic cells

Kazumasa Yoshida, Nozomi Sugimoto, Satoko Iwahori, Takashi Yugawa, Mako Narisawa-Saito, Tohru Kiyono, Masatoshi Fujita

研究成果: ジャーナルへの寄稿記事

20 引用 (Scopus)

抄録

CDC6, a replication licensing protein, is partially exported to the cytoplasm in human cells through phosphorylation by Cdk during S phase, but a significant proportion remains in the nucleus. We report here that human CDC6 physically interacts with ATR, a crucial checkpoint kinase, in a manner that is stimulated by phosphorylation by Cdk. CDC6 silencing by siRNAs affected ATR-dependent inhibition of mitotic entry elicited by modest replication stress. Whereas a Cdk-phosphorylation-mimicking CDC6 mutant could rescue the checkpoint defect by CDC6 silencing, a phosphorylation-deficient mutant could not. Furthermore, we found that the CDC6-ATR interaction is conserved in Xenopus. We show that the presence of Xenopus CDC6 during S phase is essential for Xenopus ATR to bind to chromatin in response to replication inhibition. In addition, when human CDC6 amino acid fragment 180-220, which can bind to both human and Xenopus ATR, was added to Xenopus egg extracts after assembly of the pre-replication complex, Xenopus Chk1 phosphorylation was significantly reduced without lowering replication, probably through a sequestration of CDC6-mediated ATR-chromatin interaction. Thus, CDC6 might regulate replication-checkpoint activation through the interaction with ATR in higher eukaryotic cells.

元の言語英語
ページ(範囲)225-235
ページ数11
ジャーナルJournal of cell science
123
発行部数2
DOI
出版物ステータス出版済み - 1 15 2010

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Eukaryotic Cells
Xenopus
Phosphorylation
S Phase
Chromatin
Licensure
Ovum
Cytoplasm
Phosphotransferases
Amino Acids
Proteins

All Science Journal Classification (ASJC) codes

  • Cell Biology

これを引用

CDC6 interaction with ATR regulates activation of a replication checkpoint in higher eukaryotic cells. / Yoshida, Kazumasa; Sugimoto, Nozomi; Iwahori, Satoko; Yugawa, Takashi; Narisawa-Saito, Mako; Kiyono, Tohru; Fujita, Masatoshi.

:: Journal of cell science, 巻 123, 番号 2, 15.01.2010, p. 225-235.

研究成果: ジャーナルへの寄稿記事

Yoshida, Kazumasa ; Sugimoto, Nozomi ; Iwahori, Satoko ; Yugawa, Takashi ; Narisawa-Saito, Mako ; Kiyono, Tohru ; Fujita, Masatoshi. / CDC6 interaction with ATR regulates activation of a replication checkpoint in higher eukaryotic cells. :: Journal of cell science. 2010 ; 巻 123, 番号 2. pp. 225-235.
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abstract = "CDC6, a replication licensing protein, is partially exported to the cytoplasm in human cells through phosphorylation by Cdk during S phase, but a significant proportion remains in the nucleus. We report here that human CDC6 physically interacts with ATR, a crucial checkpoint kinase, in a manner that is stimulated by phosphorylation by Cdk. CDC6 silencing by siRNAs affected ATR-dependent inhibition of mitotic entry elicited by modest replication stress. Whereas a Cdk-phosphorylation-mimicking CDC6 mutant could rescue the checkpoint defect by CDC6 silencing, a phosphorylation-deficient mutant could not. Furthermore, we found that the CDC6-ATR interaction is conserved in Xenopus. We show that the presence of Xenopus CDC6 during S phase is essential for Xenopus ATR to bind to chromatin in response to replication inhibition. In addition, when human CDC6 amino acid fragment 180-220, which can bind to both human and Xenopus ATR, was added to Xenopus egg extracts after assembly of the pre-replication complex, Xenopus Chk1 phosphorylation was significantly reduced without lowering replication, probably through a sequestration of CDC6-mediated ATR-chromatin interaction. Thus, CDC6 might regulate replication-checkpoint activation through the interaction with ATR in higher eukaryotic cells.",
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AU - Narisawa-Saito, Mako

AU - Kiyono, Tohru

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