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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: Jun 22, 2026

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51

Published on: February 13, 2019

Fbh1 limits Rad51-dependent recombination at blocked replication forks.

Alexander Lorenz¹, Fekret Osman, Victoria Folkyte

¹Department of Biochemistry, University of Oxford, Oxford, United Kingdom.

Molecular and Cellular Biology

|June 24, 2009

Summary

F-box DNA helicase (Fbh1) in fission yeast opposes Rad22 to limit Rad51 loading onto DNA, preventing recombination during replication stress. This function is crucial for DNA repair, distinct from spontaneous recombination control.

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Last Updated: Jun 22, 2026

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51

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Published on: February 13, 2019

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

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Visualization of DNA Repair Proteins Interaction by Immunofluorescence

Visualization of DNA Repair Proteins Interaction by Immunofluorescence

Published on: June 26, 2020

Area of Science:

Molecular Biology
Genetics
DNA Repair Mechanisms

Background:

Controlling Rad51 loading onto DNA is critical for regulating homologous recombination.
Fbh1 is an F-box DNA helicase with a known role in DNA repair pathways.
Rad51 is a key protein in homologous recombination, essential for DNA repair and genome stability.

Purpose of the Study:

To investigate the role of F-box DNA helicase (Fbh1) in controlling Rad51 loading and homologous recombination in fission yeast.
To determine if Fbh1's activity is involved in preventing spontaneous or replication-stress-induced recombination.
To elucidate the mechanism by which Fbh1 regulates Rad51 loading and its relationship with other recombination factors like Rad22 and Srs2.

Main Methods:

Genetic assays in fission yeast.
Indirect immunofluorescence to visualize Rad51 foci.
Analysis of recombination events under conditions of replication stress and fork barriers.
Comparative studies using Fbh1 and Srs2 mutants and overexpression strains.

Main Results:

Fbh1 directly opposes Rad22 to curb Rad51 loading onto DNA.
Fbh1's activity is essential for preventing recombination when replication forks are blocked or broken, but not for spontaneous recombination.
Overexpression of Fbh1 reduces replication fork block-induced recombination and Rad51 nuclear foci, dependent on its helicase/translocase activity.
Fbh1 acts as a Rad51 disruptase, suppressing recombination and Rad51 accumulation at replication fork barriers, a function partially overlapping but distinct from Srs2.

Conclusions:

Fbh1 plays a critical role in preventing homologous recombination during replication stress by limiting Rad51 loading.
Fbh1 functions as a Rad51 disruptase, with a distinct role compared to the known Rad51 disruptase Srs2.
Fbh1's activity is vital for maintaining genome stability under conditions of replication stress.