Functional relationship between Claspin and Rad17

Akari Yoshimura, Motomu Akita, Yoshifumi Hosono, Takuya Abe, Masahiko Kobayashi, Ken Ichi Yamamoto, Shusuke Tada, Masayuki Seki, Takemi Enomoto

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Claspin was originally identified as a Check1 (Chk1)-interacting protein. Claspin and Rad17 are reportedly involved in the DNA damage-induced phosphorylation of Chk1, a hallmark of checkpoint activation. To understand the cellular functions of Claspin and the functional relationship between Claspin and Rad17, we generated Claspin -/- and Claspin -/-/RAD17 - cells using chicken DT40 cells, which contain an exogenously introduced Claspin that can be suppressed by the addition of doxycycline (Dox). In the presence of Dox, Claspin -/- cells ceased growth within 2. days, leading to cell death. In addition, a remarkable reduction in the rate of DNA elongation was observed in Claspin-depleted cells, suggesting that Claspin plays a critical role in DNA replication in the absence of exogenous stress. When cells were exposed to methyl methanesulfonate (MMS), a DNA damaging agent, RAD17 - cells showed a greater defect in checkpoint activation than Claspin -/- cells as monitored by progression of cell cycle and phosphorylation of Chk1. Knocking out RAD17 gene showed almost no additive effects on cell death and DNA elongation rates in Claspin-depleted cells.

Original languageEnglish
Pages (from-to)298-303
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume414
Issue number2
DOIs
Publication statusPublished - Oct 22 2011
Externally publishedYes

Keywords

  • Claspin
  • DNA replication
  • DT40
  • Rad17

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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  • Cite this

    Yoshimura, A., Akita, M., Hosono, Y., Abe, T., Kobayashi, M., Yamamoto, K. I., Tada, S., Seki, M., & Enomoto, T. (2011). Functional relationship between Claspin and Rad17. Biochemical and Biophysical Research Communications, 414(2), 298-303. https://doi.org/10.1016/j.bbrc.2011.09.037