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Proofreading

Proofreading

Synthesis of new DNA molecules is carried out by the enzyme DNA polymerase, which adds nucleotides on the daughter strand complementary to the template DNA strand. DNA polymerase has a higher affinity to add the correct base and ensures fidelity during DNA replication. Furthermore, it exhibits proofreading activity during replication, using an exonuclease domain that cuts off incorrect nucleotides from the nascent DNA strand.
Errors During Replication are Corrected by the DNA Polymerase...

Proofreading

Proofreading

Replication in Eukaryotes

Replication in Eukaryotes

Replication in Eukaryotes

Replication in Eukaryotes

In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before cell division, so there are many proteins that fulfill specialized roles in the replication process. Replication occurs in three phases: initiation, elongation, and termination, and ends with two complete sets of chromosomes in the nucleus.
Many Proteins Orchestrate Replication at the Origin
Eukaryotic replication follows many of the same...

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...

The DNA Replication Fork

The DNA Replication Fork

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Related Experiment Video

Updated: Mar 9, 2026

Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method

Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method

Published on: May 2, 2025

Quality control mechanisms exclude incorrect polymerases from the eukaryotic replication fork.

Grant D Schauer¹, Michael E O'Donnell²

¹Howard Hughes Medical Institute and Rockefeller University, New York, NY 10065.

Proceedings of the National Academy of Sciences of the United States of America

|January 11, 2017

Summary

This summary is machine-generated.

Replication factor C (RFC) prevents the wrong DNA polymerase from replicating the lagging strand, while the CMG helicase protects the correct polymerase on the leading strand, ensuring genome stability.

Keywords:

DNA polymerase PCNA clamp loader replication replisome

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Published on: April 29, 2010

Area of Science:

Molecular Biology
Genetics
Biochemistry

Background:

Eukaryotic DNA replication involves DNA polymerase ε (Pol ε) and Pol δ on leading and lagging strands, respectively.
Recruitment is mediated by the CMG helicase for Pol ε and PCNA for Pol δ.
Mechanisms ensuring polymerase-lagging strand specificity were previously unclear.

Purpose of the Study:

To identify quality control mechanisms preventing incorrect polymerase assignment to DNA strands.
To elucidate the roles of RFC and CMG in polymerase fidelity during replication.

Main Methods:

Investigated polymerase binding affinities to CMG using kinetic measurements.
Assessed the inhibitory effects of RFC on Pol ε in vitro.
Analyzed polymerase behavior in the context of CMG and PCNA interactions.

Main Results:

RFC specifically inhibits Pol ε on the lagging strand.
CMG protects Pol ε from RFC inhibition on the leading strand.
Pol δ does not bind CMG, and Pol δ-PCNA is ejected from the leading strand via collision release.

Conclusions:

RFC and CMG act as crucial quality control factors, ensuring correct polymerase function on each DNA strand.
These mechanisms prevent polymerase-CMG/PCNA conflicts and maintain replication fidelity.
The study reveals a sophisticated clamp machinery for precise eukaryotic DNA replication.