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The Replisome

The Replisome

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Related Experiment Video

Updated: Oct 1, 2025

Author Spotlight: Investigating the Motion Dynamics of the Eukaryotic Replisome Components at the Single-Molecule Level

Author Spotlight: Investigating the Motion Dynamics of the Eukaryotic Replisome Components at the Single-Molecule Level

Published on: July 26, 2024

Single-molecule mapping of replisome progression.

Clémence Claussin¹, Jacob Vazquez¹, Iestyn Whitehouse¹

¹Molecular Biology Program, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, NY 10065, USA.

|March 3, 2022

Summary

New Replicon-seq technology maps DNA replication forks, revealing that sister replisomes move independently but consistently. This method tracks replication dynamics and identifies key roles for the Rrm3 helicase in overcoming obstacles.

Keywords:

DNA replication Nanopore sequencing Replicon-seq Rrm3 rDNA replication termination replisome pausing sister replisome

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Author Spotlight: Unraveling the Dynamics of Eukaryotic DNA Replication Through Single-Molecule Visualization

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Related Experiment Videos

Last Updated: Oct 1, 2025

Author Spotlight: Investigating the Motion Dynamics of the Eukaryotic Replisome Components at the Single-Molecule Level

Author Spotlight: Investigating the Motion Dynamics of the Eukaryotic Replisome Components at the Single-Molecule Level

Published on: July 26, 2024

Visualization of Replisome Encounters with an Antigen Tagged Blocking Lesion

Visualization of Replisome Encounters with an Antigen Tagged Blocking Lesion

Published on: July 27, 2021

Author Spotlight: Unraveling the Dynamics of Eukaryotic DNA Replication Through Single-Molecule Visualization

Author Spotlight: Unraveling the Dynamics of Eukaryotic DNA Replication Through Single-Molecule Visualization

Published on: September 27, 2024

Area of Science:

Molecular Biology
Genetics
Genomics

Background:

Understanding DNA replication is crucial, but precise mapping of replication machinery on chromatin is challenging.
Key questions remain regarding replication stall sites, replisome-gene transcription interactions, and sister replisome coordination.

Purpose of the Study:

To develop a novel method for high-resolution mapping of individual DNA replication sites.
To investigate the coordination and dynamics of sister replisomes during DNA replication.

Main Methods:

Developed Replicon-seq: a technique for excising full-length replicons via nuclease cleavage at replication forks.
Utilized Nanopore sequencing for single-molecule, long-read DNA analysis of excised replicons.

Main Results:

Demonstrated that sister replisomes operate autonomously yet exhibit consistent progression through chromatin.
Observed transient pausing of replication forks at obstacles, followed by rapid resumption of DNA synthesis.
Identified the helicase Rrm3 as critical for mitigating protein barriers and facilitating replication termination.

Conclusions:

Replicon-seq offers a high-resolution approach to study individual replisome movement across the genome.
Sister replisome autonomy and coordinated progression are key features of DNA replication.
The Rrm3 helicase is essential for efficient replication fork progression and termination.