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Restarting Stalled Replication Forks

Restarting Stalled Replication Forks

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...

Restarting Stalled Replication Forks

Restarting Stalled Replication Forks

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...

The DNA Replication Fork

The DNA Replication Fork

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 forks, one in...

The DNA Replication Fork

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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 forks, one in...

Translesion DNA Polymerases

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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
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DNA Replication

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DNA replication involves the separation of the two strands of the double helix, with each strand serving as a template from which the new complementary strand is copied. After replication, each double-stranded DNA includes one parental or “old” strand and one “new” strand. This is known as semiconservative replication. The resulting DNA molecules have the same sequence and are divided equally into the two daughter cells.
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Related Experiment Video

Updated: May 25, 2026

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

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

Published on: October 27, 2011

dNTP pools determine fork progression and origin usage under replication stress.

Jérôme Poli¹, Olga Tsaponina, Laure Crabbé

¹Institute of Human Genetics, CNRS UPR 1142, Montpellier, France.

The EMBO Journal

|January 12, 2012

Summary

Maintaining balanced deoxyribonucleoside triphosphate (dNTP) levels is crucial for genome stability. This study reveals how dNTP pool changes impact DNA replication dynamics and adaptation to replication stress in yeast.

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Last Updated: May 25, 2026

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

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Published on: October 27, 2011

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

Nucleoside Triphosphates - From Synthesis to Biochemical Characterization

Nucleoside Triphosphates - From Synthesis to Biochemical Characterization

Published on: April 3, 2014

Area of Science:

Molecular Biology
Genetics
Cell Biology

Background:

Intracellular deoxyribonucleoside triphosphate (dNTP) pools are vital for genome integrity.
Altered dNTP levels are linked to mutagenesis, genomic instability, and cancer.
The precise mechanisms linking dNTP pool changes to DNA synthesis are not fully understood.

Purpose of the Study:

To investigate how intracellular dNTP levels influence DNA replication dynamics in budding yeast.
To explore the role of ribonucleotide reductase (RNR) in regulating dNTP pools and replication.
To understand the adaptation mechanisms of chromosomal instability (CIN) mutants under replication stress.

Main Methods:

Utilized budding yeast as a model organism.
Manipulated intracellular dNTP levels by altering ribonucleotide reductase (RNR) activity.
Applied hydroxyurea (HU) to induce replication stress.
Observed effects on DNA replication fork speed, origin usage, and DNA synthesis.

Main Results:

Increased dNTP pools, via RNR upregulation, enhance DNA replication elongation, suggesting dNTPs are normally limiting.
Hydroxyurea (HU) treatment rapidly shifts DNA replication to a slower mode.
RNR upregulation delays this shift and modulates fork speed and origin usage under HU-induced stress.
Chromosomal instability (CIN) mutants exhibit elevated dNTP pools and improved DNA synthesis during HU treatment.

Conclusions:

Intracellular dNTP levels significantly impact DNA replication dynamics.
CIN mutants may adapt to chronic replication stress by upregulating dNTP pools via RNR.
This adaptation potentially enhances fork progression in the presence of DNA damage.