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Related Concept Videos

The Replisome03:01

The Replisome

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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Dynamics of pre-replicative complex assembly.

Vasiliki Tsakraklides1, Stephen P Bell2

  • 1Department of Biology, Cambridge, Massachusetts 02139.

The Journal of Biological Chemistry
|January 26, 2010
PubMed
Summary
This summary is machine-generated.

Pre-replicative complex (pre-RC) assembly involves sequential binding events. Helicase loading during pre-RC formation requires ATP hydrolysis and results in the dynamic dissociation of key proteins from origin DNA.

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Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
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Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method

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Area of Science:

  • Molecular Biology
  • Cell Cycle Regulation
  • DNA Replication

Background:

  • The pre-replicative complex (pre-RC) is essential for initiating DNA replication.
  • Formation of the pre-RC involves the Origin Recognition Complex (ORC), Cdc6, Cdt1, and the Mcm2-7 complex.
  • The Mcm2-7 complex functions as the replicative helicase, loaded onto origin DNA.

Purpose of the Study:

  • To investigate the step-by-step assembly of the pre-RC in vitro.
  • To determine the dynamic behavior of pre-RC components during helicase loading.
  • To elucidate the role of ATP hydrolysis in pre-RC formation.

Main Methods:

  • In vitro pre-RC assembly assays.
  • Stepwise analysis of protein-DNA interactions.
  • Investigation of protein dissociation during helicase loading.

Main Results:

  • Pre-RC formation occurs through distinct, separable binding events.
  • Origin Recognition Complex (ORC) recruits Cdc6, forming a ternary complex.
  • Helicase loading is coupled with the ATP-dependent dissociation of ORC, Cdc6, and Cdt1 from origin DNA.

Conclusions:

  • Pre-RC assembly is a dynamic process, not a static complex formation.
  • ATP hydrolysis is crucial for the dissociation of components during the late stages of pre-RC assembly.
  • Understanding these dynamics provides insights into the regulation of DNA replication initiation.