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Maturation-associated Dbf4 expression is essential for mouse zygotic DNA replication.

Shin Murai1, Yukiko Katagiri, Shigeru Yamashita

  • 1Department of Biochemistry, Toho University School of Medicine, 5-21-16 Omorinishi Otaku, 143-8540, Tokyo, Japan.

Development, Growth & Differentiation
|October 29, 2014
PubMed
Summary

Cdc7 kinase activity is regulated by its Dbf4 subunit during mouse oocyte maturation. Dbf4 accumulation activates Cdc7, enabling DNA replication after fertilization.

Keywords:
Cdc7/Dbf4 kinaseDNA replicationcytoplasmic polyadenylationmouse zygoteoocyte maturation

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Developmental Biology

Background:

  • Cdc7 is an S-phase-promoting kinase essential for DNA replication initiation.
  • Cdc7 phosphorylates the MCM complex, which functions as the replicative helicase.
  • Cdc7 activity is absent in immature mouse oocytes but present in mature oocytes and early embryos.

Purpose of the Study:

  • To investigate the regulation of Cdc7 kinase activity during mouse oocyte maturation.
  • To identify the role of the Dbf4 regulatory subunit in Cdc7 activation.
  • To understand the mechanism controlling Cdc7 activity before zygotic DNA replication.

Main Methods:

  • RNA interference (RNAi) to knockdown maternal Dbf4 expression.
  • Western blotting to detect protein levels of Dbf4 and Cdc7.
  • Analysis of DNA replication in one-cell embryos after Dbf4 knockdown.

Main Results:

  • Dbf4 protein accumulates during oocyte maturation, correlating with Cdc7 activation.
  • RNAi-mediated Dbf4 knockdown prevented Cdc7 activation and DNA replication.
  • Dbf4 mRNA undergoes cytoplasmic polyadenylation, dependent on MPF activity, during maturation.

Conclusions:

  • Dbf4 is the key regulator of Cdc7 activity in mouse oocytes.
  • Cdc7 is inactivated in immature oocytes to prevent premature MCM phosphorylation.
  • Oocyte maturation is crucial for activating Cdc7 and enabling subsequent embryonic development.