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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

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...

Homologous Recombination

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...

Homologous Recombination

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...

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Related Experiment Video

Updated: May 23, 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

Replication fork reversal after replication-transcription collision.

Anne L De Septenville¹, Stéphane Duigou, Hasna Boubakri

¹CNRS, Centre de Génétique Moléculaire, UPR3404, Gif-sur-Yvette, France.

|April 13, 2012

Summary

Recombination proteins are crucial for repairing DNA breaks caused by replication-transcription collisions in E. coli. RecBC is essential for viability, preventing lethal fork breakage and degradation.

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Inducing a Site Specific Replication Blockage in E. coli Using a Fluorescent Repressor Operator System

Inducing a Site Specific Replication Blockage in E. coli Using a Fluorescent Repressor Operator System

Published on: August 21, 2016

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Last Updated: May 23, 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

Inducing a Site Specific Replication Blockage in E. coli Using a Fluorescent Repressor Operator System

Inducing a Site Specific Replication Blockage in E. coli Using a Fluorescent Repressor Operator System

Published on: August 21, 2016

Area of Science:

Molecular Biology
Genetics
Microbiology

Background:

Replication fork arrest is a significant source of genetic instability.
Transcription-replication collisions are a major cause of replication impediment.

Purpose of the Study:

To investigate the role of recombination proteins in Escherichia coli during replication-transcription head-on collisions.
To identify essential recombination proteins for cell viability under collision conditions.

Main Methods:

Inducing replication-transcription head-on collisions via ribosomal operon inversion in E. coli.
Analyzing the requirement for recombination proteins (RecBC, RecA, RecD, RuvABC) and exonucleases (RecJ) in cell viability.
Characterizing DNA breakage and degradation in mutant strains.

Main Results:

RecBC is the sole recombination protein essential for viability during induced collisions.
Absence of RecBC leads to unrepaired fork breakage and degradation by RecJ exonuclease.
RecA and RecD inactivation causes lethal fork breakage, with degradation by RecBC helicase and RecJ exonuclease.
DNA degradation is hindered by proximity to another rrn operon.
Holliday junction resolution by RuvABC contributes to linear DNA formation in recB and recA recD mutants.

Conclusions:

Replication fork reversal is proposed to occur post-collision, facilitating accessory helicase action to remove obstacles.
RecBC plays a critical role in resolving replication-transcription conflicts and maintaining genome stability.