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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.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
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.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
S-Cdk Initiates DNA Replication02:38

S-Cdk Initiates DNA Replication

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).
Two states at the origin of replication
In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called the origins of replication.
S-Cdk Initiates DNA Replication02:38

S-Cdk Initiates DNA Replication

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).
Two states at the origin of replication
In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called the origins of replication.
Replication in Eukaryotes01:29

Replication in Eukaryotes

In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before cell division, so there are many proteins that fulfill specialized roles in the replication process. Replication occurs in three phases: initiation, elongation, and termination, and ends with two complete sets of chromosomes in the nucleus.
Many Proteins Orchestrate Replication at the Origin
Eukaryotic replication follows many of the same...
Replication in Eukaryotes02:31

Replication in Eukaryotes

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Related Experiment Video

Updated: May 21, 2026

Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
08:53

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

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Dynamic association of ORCA with prereplicative complex components regulates DNA replication initiation.

Zhen Shen1, Arindam Chakraborty, Ankur Jain

  • 1Department of Cell and Developmental Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.

Molecular and Cellular Biology
|May 31, 2012
PubMed
Summary

Origin recognition complex-associated (ORCA) protein is essential for DNA replication initiation in human cells. ORCA dynamically binds to replication proteins, regulating their assembly and facilitating the G(1)-to-S-phase transition.

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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:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • DNA replication initiation in eukaryotes depends on prereplicative complex (pre-RC) assembly at origins.
  • Origin recognition complex (ORC)-associated (ORCA/LRWD1) protein stabilizes ORC binding to chromatin.

Purpose of the Study:

  • To investigate the role of ORCA in the G(1)-to-S-phase transition and its interactions with pre-RC components.
  • To elucidate the dynamic binding of ORCA with ORC, Cdt1, and geminin throughout the cell cycle.

Main Methods:

  • Cell cycle analysis in human cells.
  • Protein-protein interaction studies using pulldown assays.
  • Single-molecule pulldown experiments.

Main Results:

  • ORCA is crucial for the G(1)-to-S-phase transition.
  • ORCA binds to ORC, Cdt1, and geminin in a specific stoichiometry.
  • ORCA's stability depends on Orc2 interaction; its association with Cdt1 and geminin varies across the cell cycle.
  • Geminin overexpression disrupts ORCA-Cdt1 interaction, suggesting titration by geminin in post-G(1) cells.

Conclusions:

  • ORCA plays a key role in regulating DNA replication initiation.
  • The dynamic association of ORCA with pre-RC components modulates protein assembly on chromatin.
  • ORCA facilitates DNA replication initiation through dynamic interactions with ORC, Cdt1, and geminin.