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Real‐Time Fluorescence Imaging of the DNA Damage Repair Response During Mitosis
Author(s) -
Miwa Shinji,
Yano Shuya,
Yamamoto Mako,
Matsumoto Yasunori,
Uehara Fuminari,
Hiroshima Yukihiko,
Toneri Makoto,
Murakami Takashi,
Kimura Hiroaki,
Hayashi Katsuhiro,
Yamamoto Norio,
Efimova Elena V.,
Tsuchiya Hiroyuki,
Hoffman Robert M.
Publication year - 2015
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.25021
Subject(s) - mitosis , mitotic catastrophe , green fluorescent protein , microbiology and biotechnology , binucleated cells , dna damage , apoptosis , chemistry , confocal microscopy , fluorescence microscope , cell cycle , biology , dna , fluorescence , biochemistry , gene , organic chemistry , toxicity , micronucleus , micronucleus test , physics , quantum mechanics
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. © 2014 Wiley Periodicals, Inc.