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Related Concept Videos

Proofreading01:31

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 Enzyme
Proofreading01:43

Proofreading

Synthesis of new DNA molecules starts when DNA polymerase links nucleotides together in a sequence that is complementary to the template DNA strand. DNA polymerase has a higher affinity for the correct base to ensure fidelity in DNA replication. The DNA polymerase furthermore proofreads 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 EnzymeGenomic DNA is synthesized in...
The Replisome03:01

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...
Translesion DNA Polymerases02:10

Translesion DNA Polymerases

Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
Replication in Prokaryotes01:32

Replication in Prokaryotes

DNA replication has three main steps: initiation, elongation, and termination. Replication in prokaryotes begins when initiator proteins bind to the single origin of replication (ori) on the cell's circular chromosome. Replication then proceeds around the entire circle of the chromosome in each direction from the two replication forks, resulting in two DNA molecules.
Many Proteins Work Together to Replicate the Chromosome
Replication is coordinated and carried out by a host of specialized...
Replication in Prokaryotes02:35

Replication in Prokaryotes

Overview

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Related Experiment Video

Updated: Jun 3, 2026

Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes
05:37

Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes

Published on: April 4, 2025

DNA polymerase activity at the single-molecule level.

Joshua P Gill1, Jun Wang, David P Millar

  • 1Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.

Biochemical Society Transactions
|March 25, 2011
PubMed
Summary
This summary is machine-generated.

DNA polymerases ensure accurate DNA replication and repair. Single-molecule fluorescence spectroscopy reveals their dynamic conformational changes during nucleotide selection and proofreading.

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Last Updated: Jun 3, 2026

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Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay
17:03

Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay

Published on: March 23, 2010

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • DNA polymerases are crucial enzymes for DNA replication and repair in all living organisms.
  • High fidelity DNA replication requires precise selection of nucleotide substrates based on the template strand.
  • Proofreading, involving a 3'→5' exonuclease activity, corrects misincorporated nucleotides.

Purpose of the Study:

  • To investigate the dynamic conformational changes in DNA polymerases during nucleotide incorporation and proofreading.
  • To utilize single-molecule fluorescence spectroscopy to observe these enzyme dynamics in real-time.

Main Methods:

  • Single-molecule fluorescence spectroscopy was employed to monitor DNA polymerase activity.
  • The study focused on observing conformational rearrangements of the polymerase-DNA complex without population synchronization.

Main Results:

  • Large conformational rearrangements were observed during both nucleotide incorporation and proofreading steps.
  • Real-time observation of these dynamic processes was achieved using single-molecule techniques.

Conclusions:

  • Single-molecule fluorescence spectroscopy is a powerful tool for studying the real-time dynamics of DNA polymerases.
  • Understanding these conformational changes is key to elucidating the mechanisms of DNA replication fidelity and proofreading.