Real-Time Fluorescence Imaging of the DNA Damage Repair Response during Mitosis

Shinji Miwa; Shuya Yano; Mako Yamamoto; Yasunori Matsumoto; Fuminari Uehara; Yukihiko Hiroshima; Makoto Toneri; Takashi Murakami; Hiroaki Kimura; Katsuhiro Hayashi; Norio Yamamoto; Elena V. Efimova; Hiroyuki Tsuchiya; Robert M. Hoffman

doi:10.1002/jcb.25021

Real-Time Fluorescence Imaging of the DNA Damage Repair Response during Mitosis

Shinji Miwa, Shuya Yano, Mako Yamamoto, Yasunori Matsumoto, Fuminari Uehara, Yukihiko Hiroshima, Makoto Toneri, Takashi Murakami, Hiroaki Kimura, Katsuhiro Hayashi, Norio Yamamoto, Elena V. Efimova, Hiroyuki Tsuchiya, Robert M. Hoffman

Research output: Contribution to journal › Article › peer-review

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Abstract

The response to DNA damage during mitosis was visualized using real-time fluorescence imaging of focus formation by the DNA-damage repair (DDR) response protein 53BP1 linked to green fluorescent protein (GFP) (53BP1-GFP) in the MiaPaCa-2^Tet-On pancreatic cancer cell line. To observe 53BP1-GFP foci during mitosis, MiaPaCa-2^Tet-On 53BP1-GFP cells were imaged every 30 min by confocal microscopy. Time-lapse imaging demonstrated that 11.4 ± 2.1% of the mitotic MiaPaCa-2^Tet-On 53BP1-GFP cells had increased focus formation over time. Non-mitotic cells did not have an increase in 53BP1-GFP focus formation over time. Some of the mitotic MiaPaCa-2^Tet-On 53BP1-GFP cells with focus formation became apoptotic. The results of the present report suggest that DNA strand breaks occur during mitosis and undergo repair, which may cause some of the mitotic cells to enter apoptosis in a phenomenon possibly related to mitotic catastrophe. J. Cell. Biochem. 116: 661-666, 2015.

Original language	English
Pages (from-to)	661-666
Number of pages	6
Journal	Journal of Cellular Biochemistry
Volume	116
Issue number	4
DOIs	https://doi.org/10.1002/jcb.25021
Publication status	Published - Apr 1 2015
Externally published	Yes

Keywords

53BP1
APOPTOSIS
DNA DAMAGE
DNA REPAIR RESPONSE
FOCUS FORMATION
FUSION GENE
GFP
MITOSIS
MITOTIC CATASTROPHE
TIME-LAPSE CONFOCAL IMAGING

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Cell Biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1002/jcb.25021

Cite this

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title = "Real-Time Fluorescence Imaging of the DNA Damage Repair Response during Mitosis",

abstract = "The response to DNA damage during mitosis was visualized using real-time fluorescence imaging of focus formation by the DNA-damage repair (DDR) response protein 53BP1 linked to green fluorescent protein (GFP) (53BP1-GFP) in the MiaPaCa-2Tet-On pancreatic cancer cell line. To observe 53BP1-GFP foci during mitosis, MiaPaCa-2Tet-On 53BP1-GFP cells were imaged every 30 min by confocal microscopy. Time-lapse imaging demonstrated that 11.4 ± 2.1% of the mitotic MiaPaCa-2Tet-On 53BP1-GFP cells had increased focus formation over time. Non-mitotic cells did not have an increase in 53BP1-GFP focus formation over time. Some of the mitotic MiaPaCa-2Tet-On 53BP1-GFP cells with focus formation became apoptotic. The results of the present report suggest that DNA strand breaks occur during mitosis and undergo repair, which may cause some of the mitotic cells to enter apoptosis in a phenomenon possibly related to mitotic catastrophe. J. Cell. Biochem. 116: 661-666, 2015.",

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author = "Shinji Miwa and Shuya Yano and Mako Yamamoto and Yasunori Matsumoto and Fuminari Uehara and Yukihiko Hiroshima and Makoto Toneri and Takashi Murakami and Hiroaki Kimura and Katsuhiro Hayashi and Norio Yamamoto and Efimova, {Elena V.} and Hiroyuki Tsuchiya and Hoffman, {Robert M.}",

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doi = "10.1002/jcb.25021",

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volume = "116",

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T1 - Real-Time Fluorescence Imaging of the DNA Damage Repair Response during Mitosis

AU - Miwa, Shinji

AU - Yano, Shuya

AU - Yamamoto, Mako

AU - Matsumoto, Yasunori

AU - Uehara, Fuminari

AU - Hiroshima, Yukihiko

AU - Toneri, Makoto

AU - Murakami, Takashi

AU - Kimura, Hiroaki

AU - Hayashi, Katsuhiro

AU - Yamamoto, Norio

AU - Efimova, Elena V.

AU - Tsuchiya, Hiroyuki

AU - Hoffman, Robert M.

PY - 2015/4/1

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N2 - The response to DNA damage during mitosis was visualized using real-time fluorescence imaging of focus formation by the DNA-damage repair (DDR) response protein 53BP1 linked to green fluorescent protein (GFP) (53BP1-GFP) in the MiaPaCa-2Tet-On pancreatic cancer cell line. To observe 53BP1-GFP foci during mitosis, MiaPaCa-2Tet-On 53BP1-GFP cells were imaged every 30 min by confocal microscopy. Time-lapse imaging demonstrated that 11.4 ± 2.1% of the mitotic MiaPaCa-2Tet-On 53BP1-GFP cells had increased focus formation over time. Non-mitotic cells did not have an increase in 53BP1-GFP focus formation over time. Some of the mitotic MiaPaCa-2Tet-On 53BP1-GFP cells with focus formation became apoptotic. The results of the present report suggest that DNA strand breaks occur during mitosis and undergo repair, which may cause some of the mitotic cells to enter apoptosis in a phenomenon possibly related to mitotic catastrophe. J. Cell. Biochem. 116: 661-666, 2015.

AB - The response to DNA damage during mitosis was visualized using real-time fluorescence imaging of focus formation by the DNA-damage repair (DDR) response protein 53BP1 linked to green fluorescent protein (GFP) (53BP1-GFP) in the MiaPaCa-2Tet-On pancreatic cancer cell line. To observe 53BP1-GFP foci during mitosis, MiaPaCa-2Tet-On 53BP1-GFP cells were imaged every 30 min by confocal microscopy. Time-lapse imaging demonstrated that 11.4 ± 2.1% of the mitotic MiaPaCa-2Tet-On 53BP1-GFP cells had increased focus formation over time. Non-mitotic cells did not have an increase in 53BP1-GFP focus formation over time. Some of the mitotic MiaPaCa-2Tet-On 53BP1-GFP cells with focus formation became apoptotic. The results of the present report suggest that DNA strand breaks occur during mitosis and undergo repair, which may cause some of the mitotic cells to enter apoptosis in a phenomenon possibly related to mitotic catastrophe. J. Cell. Biochem. 116: 661-666, 2015.

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KW - MITOTIC CATASTROPHE

KW - TIME-LAPSE CONFOCAL IMAGING

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ER -

Real-Time Fluorescence Imaging of the DNA Damage Repair Response during Mitosis

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

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