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Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart, a...
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In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
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Aurora-A controls pre-replicative complex assembly and DNA replication by stabilizing geminin in mitosis.

Takaaki Tsunematsu1, Yoshihiro Takihara, Naozumi Ishimaru

  • 1Department of Oral and Maxillofacial Pathobiology, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan.

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|May 23, 2013
PubMed
Summary

Geminin controls DNA replication by binding Cdt1. Aurora-A stabilizes geminin during mitosis, preventing Cdt1 degradation and ensuring proper DNA replication in the next cell cycle.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • Geminin is crucial for DNA replication, inhibiting re-replication by binding Cdt1.
  • Geminin levels are regulated by the anaphase-promoting complex/cyclosome (APC/C) during the cell cycle.
  • Mitotic regulation of geminin stability and its role in subsequent DNA replication were previously undescribed.

Purpose of the Study:

  • To elucidate the role of geminin regulation during M phase.
  • To investigate the mechanism of geminin stabilization during mitosis.
  • To understand how geminin influences Cdt1 stability and pre-replicative complex formation in the subsequent S phase.

Main Methods:

  • Phosphorylation site mapping of geminin during M phase.
  • Analysis of geminin stability in response to Aurora-A activity.
  • Investigation of the interaction between geminin, Cdt1, and the SCF(Skp2) complex.

Main Results:

  • Aurora-A phosphorylates geminin at Thr25 during M phase.
  • This phosphorylation stabilizes geminin by preventing its degradation by the APC/C.
  • Stabilized geminin inhibits SCF(Skp2)-mediated Cdt1 degradation, promoting pre-replicative complex formation in the next S phase.

Conclusions:

  • The Aurora-A-geminin-Cdt1 axis is a critical regulator of DNA replication.
  • Geminin stabilization during mitosis by Aurora-A is essential for timely and accurate DNA replication.
  • This pathway ensures the proper formation of pre-replicative complexes for the subsequent S phase.