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S-Cdk Initiates DNA Replication02:38

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The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
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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,...
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An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork.   Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication...
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DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
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Updated: Aug 17, 2025

Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
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DDK promotes DNA replication initiation: Mechanistic and structural insights.

Ningning Li1, Ning Gao2, Yuanliang Zhai3

  • 1State Key Laboratory of Membrane Biology, Peking-Tsinghua Joint Center for Life Sciences, School of Life Sciences, Peking University, Beijing, China.

Current Opinion in Structural Biology
|December 16, 2022
PubMed
Summary

DNA replication initiation requires careful regulation. New structural studies reveal how Dbf4-dependent kinase (DDK) activates the minichromosome maintenance (MCM) double-hexamer (DH) complex, crucial for DNA unwinding and cell cycle progression.

Keywords:
Cryo-electron microscopyDDKDNA replication initiationDbf4-dependent kinaseMCMReplicative helicase activation

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • DNA replication initiation in eukaryotes is a tightly regulated, cell-cycle-dependent process.
  • It involves replication licensing to install inactive minichromosome maintenance (MCM) double-hexamers (DH) and origin firing to assemble active Cdc45-Mcm2-7-GINS (CMG) helicases.
  • Cyclin-dependent kinase (CDK) and Dbf4-dependent kinase (DDK) are key kinases regulating replication factor association with MCM-DH to form CMG helicases.

Purpose of the Study:

  • To elucidate the molecular mechanisms underlying the activation of DNA replication initiation machinery.
  • To understand the role of Dbf4-dependent kinase (DDK) in activating the minichromosome maintenance (MCM) double-hexamer (DH) complex.
  • To provide structural insights into kinase activation, substrate recognition, and phosphorylation in helicase activation.

Main Methods:

  • Cryo-electron microscopy (cryo-EM) was employed to capture structural snapshots of replication initiation complexes.
  • Structural characterization of the minichromosome maintenance (MCM) double-hexamer (DH) in complex with Dbf4-dependent kinase (DDK).

Main Results:

  • Recent cryo-EM studies have provided a framework for understanding replication initiation machinery assembly and regulation.
  • The structural characterization of the MCM-DH-DDK complex offers insights into DDK-mediated MCM-DH phosphorylation.
  • This phosphorylation is critical for activating the MCM helicase for DNA unwinding.

Conclusions:

  • The structural elucidation of the MCM-DH-DDK complex significantly advances our understanding of eukaryotic DNA replication initiation.
  • DDK plays a pivotal role in helicase activation through specific phosphorylation of the MCM-DH complex.
  • These findings provide a molecular basis for understanding the regulation of DNA replication initiation and its fidelity.