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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...

Single-Strand DNA Binding Proteins

Single-Strand DNA Binding Proteins

For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...

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Related Experiment Video

Updated: Jun 28, 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

RecG interacts directly with SSB: implications for stalled replication fork regression.

Jackson A Buss¹, Yuji Kimura, Piero R Bianco

¹Department of Microbiology and Immunology, Center for Single Molecule Biophysics, University at Buffalo, Buffalo, NY 14214, USA.

Nucleic Acids Research

|November 7, 2008

Summary

This summary is machine-generated.

RecG protein binds to SSB, stabilizing its interaction with single-stranded DNA, while RuvAB does not. This differential binding influences their roles in DNA replication fork repair.

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Last Updated: Jun 28, 2026

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Area of Science:

Molecular Biology
DNA Replication and Repair

Background:

RecG and RuvAB are key proteins involved in DNA replication fork restart.
Their precise roles in fork regression and the order of their action remain unclear.

Purpose of the Study:

To elucidate the distinct functions of RecG and RuvAB at stalled DNA replication forks.
To determine the mechanism by which these proteins interact with DNA structures and accessory proteins.

Main Methods:

Biochemical assays were employed to study protein-DNA interactions.
Enzyme kinetics were used to analyze the ATPase activity of RecG and RuvAB with different DNA substrates.
Binding assays assessed the interaction between RecG, RuvAB, and single-stranded DNA binding protein (SSB).

Main Results:

RecG forms a stable complex with SSB, enhancing its affinity for single-stranded DNA (ssDNA).
RuvAB does not bind to SSB.
RecG's ATPase activity is stimulated by ssDNA and negatively supercoiled DNA, whereas RuvAB's is stimulated by relaxed circular DNA.

Conclusions:

The DNA substrate available at stalled forks dictates the access of repair proteins.
RecG likely acts earlier in the fork regression pathway due to its interaction with SSB and stimulation by ssDNA.
RuvAB may act subsequently or independently, potentially utilizing different DNA structures.