Journal Highlight: Real-time fluorescence imaging of the DNA damage repair response during mitosis

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  • Published: May 11, 2015
  • Author: spectroscopyNOW
  • Channels: UV/Vis Spectroscopy
thumbnail image: Journal Highlight: Real-time fluorescence imaging of the DNA damage repair response during mitosis
The response to DNA damage during mitosis was visualized using real-time fluorescence imaging of focus formation by the DNA-damage repair response protein 53BP1 linked to green fluorescent protein in the MiaPaCa-2Tet-On pancreatic cancer cell line.

Real-time fluorescence imaging of the DNA damage repair response during mitosis

Journal of Cellular Biochemistry, 2015, 116, 661-666
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 and Robert M. Hoffman

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.

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