Related Experiment Video

Updated: Sep 22, 2025

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
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Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay

Published on: May 3, 2018

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The structural basis of Cdc7-Dbf4 kinase dependent targeting and phosphorylation of the MCM2-7 double hexamer

Almutasem Saleh1, Yasunori Noguchi1, Ricardo Aramayo1

  • 1DNA Replication Group, Institute of Clinical Sciences, Faculty of Medicine, Imperial College London, Du Cane Road, London, W12 0NN, UK.

Nature Communications
|May 25, 2022
PubMed

Insights

The Dbf4-dependent Cdc7 kinase (DDK) controls DNA replication fork assembly by phosphorylating the MCM2-7 helicase. This study reveals how DDK binds and targets MCM subunits, offering insights for new drug development.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Replication fork control is vital for genome stability.
  • The Dbf4-dependent Cdc7 kinase (DDK) initiates DNA replication by phosphorylating the MCM2-7 helicase.
  • Mechanisms of DDK docking and substrate targeting remain unclear.

Purpose of the Study:

  • To elucidate the structural basis of DDK interaction with the MCM2-7 helicase.
  • To understand how DDK selectively phosphorylates MCM subunits.
  • To identify potential targets for DDK inhibitor development.

Main Methods:

  • Cryo-electron microscopy
  • Biochemical analysis

Main Results:

  • Identified an interaction between Dbf4's HBRCT domain and Mcm2 as a key anchoring point.
  • Demonstrated DDK binding across the MCM2-7 double-hexamer interface.
  • Showed DDK phosphorylation of Mcm4, Mcm2, and Mcm6 through binding and rotation.

Conclusions:

  • Provided fundamental insights into DDK structure and MCM2-7 helicase activation.
  • Revealed the mechanism of DDK targeting and phosphorylation of MCM subunits.
  • Established a basis for developing novel DDK inhibitors for therapeutic applications.

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