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The Replisome

The Replisome

DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...

The Replisome

The Replisome

DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...

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...

S-Cdk Initiates DNA Replication

S-Cdk Initiates DNA Replication

The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
Two states at the origin of replication
In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called the origins of replication.

S-Cdk Initiates DNA Replication

S-Cdk Initiates DNA Replication

The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
Two states at the origin of replication
In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called the origins of replication.

Replication in Eukaryotes

Replication in Eukaryotes

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Hybrid Ensemble and Single-molecule Assay to Image the Motion of Fully Reconstituted CMG

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The Elg1 replication factor C-like complex functions in PCNA unloading during DNA replication.

Takashi Kubota¹, Kohei Nishimura, Masato T Kanemaki

¹Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, Scotland, UK.

|March 19, 2013

Summary

This summary is machine-generated.

The Elg1 replication factor C-like complex (Elg1-RLC) unloads the sliding clamp PCNA from DNA during replication. This process is crucial for maintaining genome stability and preventing PCNA accumulation.

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Published on: September 27, 2024

Area of Science:

Molecular Biology
Genetics
Cell Biology

Background:

Proliferating Cell Nuclear Antigen (PCNA) is a ring-shaped complex essential for DNA replication, acting as a polymerase clamp and scaffold.
PCNA is loaded onto DNA by Replication Factor C (RFC), but its removal mechanism remained unclear.

Purpose of the Study:

To investigate the mechanism of PCNA removal from DNA after Okazaki fragment completion or replication fork termination.
To identify the protein complex responsible for PCNA unloading during DNA replication.

Main Methods:

Utilized an improved degron system in Saccharomyces cerevisiae to deplete Elg1.
Performed in vitro chromatin binding assays with purified Elg1-RLC.
Analyzed PCNA localization on chromatin in wild-type and elg1Δ mutant yeast strains.

Main Results:

Depletion of Elg1 led to PCNA accumulation on Saccharomyces cerevisiae chromatin during replication.
Reintroducing Elg1 expression reversed PCNA accumulation in vivo.
Purified Elg1-RLC efficiently unloaded PCNA from chromatin in vitro.
Elg1-RLC unloads both unmodified and SUMOylated PCNA.

Conclusions:

The Elg1 replication factor C-like complex (Elg1-RLC) is responsible for unloading PCNA from DNA during replication.
Timely PCNA unloading by Elg1-RLC is critical for maintaining genome stability.
Loss of Elg1 function results in genome instability due to persistent PCNA.