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Restarting Stalled Replication Forks

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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, a...

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Related Experiment Video

Updated: May 19, 2026

Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase

Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase

Published on: April 29, 2010

Human CST promotes telomere duplex replication and general replication restart after fork stalling.

Jason A Stewart¹, Feng Wang, Mary F Chaiken

¹Department of Cancer and Cell Biology, University of Cincinnati, Cincinnati, OH 45267, USA.

The EMBO Journal

|August 7, 2012

Summary

This summary is machine-generated.

The mammalian CST complex (CTC1-STN1-TEN1) is crucial for DNA replication, particularly at telomeres and during replication stress. Depleting CST subunits impairs replication fork progression and origin firing, highlighting its role in genomic stability.

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Last Updated: May 19, 2026

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Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

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Detection of DNA Breaks in Dividing Human Cells by Neutral Comet Assay

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Published on: August 23, 2024

Area of Science:

Molecular Biology
Genetics
Cell Biology

Background:

The mammalian CST complex, comprising CTC1, STN1, and TEN1, is known to associate with telomeres.
Previous studies indicate that depletion of CTC1 or STN1 leads to telomere defects.
The precise function of the mammalian CST complex in DNA replication and genome stability remains incompletely understood.

Purpose of the Study:

To elucidate the function of the mammalian CST complex in DNA replication and genome stability.
To investigate the role of CST in overcoming replication barriers and responding to replication stress.

Main Methods:

Stable knockdown of CST subunits (CTC1, STN1).
Analysis of telomere length and genomic instability markers (anaphase bridges, multi-telomeric signals).
Assessment of DNA replication dynamics using EdU incorporation after hydroxyurea (HU) treatment to induce replication fork stalling.

Main Results:

Depletion of CTC1 or STN1 causes both telomeric and non-telomeric phenotypes, indicative of DNA replication defects.
Knockdown of CTC1 or STN1 increases genomic and telomeric instability.
STN1 depletion delays replication through telomeres and impairs genome-wide replication restart after HU-induced stalling by reducing new origin firing, rather than affecting fork restart directly.

Conclusions:

The CST complex plays a critical role in DNA replication, extending beyond telomeres.
CST is essential for facilitating replication fork progression through natural barriers and rescuing stalled forks during replication stress.
CST likely promotes genomic stability by facilitating dormant origin firing under replication stress conditions.