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 journalArticle

1 Citation (Scopus)

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.

Original languageEnglish
Pages (from-to)661-666
Number of pages6
JournalJournal of Cellular Biochemistry
Volume116
Issue number4
DOIs
Publication statusPublished - Apr 1 2015
Externally publishedYes

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

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  • Cite this

    Miwa, S., Yano, S., Yamamoto, M., Matsumoto, Y., Uehara, F., Hiroshima, Y., Toneri, M., Murakami, T., Kimura, H., Hayashi, K., Yamamoto, N., Efimova, E. V., Tsuchiya, H., & Hoffman, R. M. (2015). Real-Time Fluorescence Imaging of the DNA Damage Repair Response during Mitosis. Journal of Cellular Biochemistry, 116(4), 661-666. https://doi.org/10.1002/jcb.25021