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CRISPR-Mediated Reorganization of Chromatin Loop Structure
09:20

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Published on: September 14, 2018

Functional relationship between Claspin and Rad17.

Akari Yoshimura1, Motomu Akita, Yoshifumi Hosono

  • 1Molecular Cell Biology Laboratory, Research Institute of Pharmaceutical Sciences, Faculty of Pharmacy, Musashino University, 1-1-20 Shinmachi, Nishitokyo-shi, Tokyo 202-8585, Japan.

Biochemical and Biophysical Research Communications
|September 28, 2011
PubMed
Summary
This summary is machine-generated.

Claspin is essential for DNA replication and cell survival, playing a critical role in DNA damage response. Its absence leads to cell death and impaired DNA elongation, even without external stress.

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Published on: January 26, 2011

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • Claspin interacts with Checkpoint kinase 1 (Chk1).
  • Claspin and Rad17 are involved in DNA damage-induced Chk1 phosphorylation, crucial for checkpoint activation.
  • The precise cellular functions of Claspin and its relationship with Rad17 require further investigation.

Purpose of the Study:

  • To elucidate the cellular functions of Claspin.
  • To understand the functional relationship between Claspin and Rad17.
  • To investigate the role of Claspin in DNA replication and DNA damage response.

Main Methods:

  • Generation of Claspin-knockout (Claspin(-/-)) and Claspin/RAD17-double-knockout (Claspin(-/-)/RAD17(-)) chicken DT40 cell lines.
  • Utilized doxycycline (Dox) to suppress exogenous Claspin expression.
  • Assessed cell viability, DNA elongation rates, cell cycle progression, and Chk1 phosphorylation following methyl methanesulfonate (MMS) exposure.

Main Results:

  • Claspin depletion in DT40 cells resulted in growth arrest and cell death within 2 days, independent of exogenous stress.
  • Claspin deficiency significantly reduced DNA elongation rates, indicating a critical role in DNA replication.
  • RAD17-deficient cells exhibited a more pronounced defect in checkpoint activation upon MMS treatment compared to Claspin-deficient cells.
  • Deletion of RAD17 showed no additive effects on cell death or DNA elongation in Claspin-depleted cells.

Conclusions:

  • Claspin is indispensable for DNA replication and cell survival, even in the absence of DNA damage.
  • While both Claspin and Rad17 are involved in DNA damage response, Claspin plays a more fundamental role in DNA replication.
  • Rad17 is not essential for the cellular response to Claspin depletion.