Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Video

Updated: May 10, 2025

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

267

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

Nicholas Kusi-Appauh1, Stefan H Mueller1, Stephen F Ralph1

  • 1Molecular Horizons and School of Science, University of Wollongong.

Journal of Visualized Experiments : Jove
|April 21, 2025
PubMed
Summary

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Dpb11 facilitates the colocalization of Mec1-Ddc2 with its activators on gapped DNA.

Cell reports·2026
Same author

Direct visualisation of post-replication gap formation at the bacterial RRS.

bioRxiv : the preprint server for biology·2026
Same author

Structure of the pre-initiation complex explains CMGE biogenesis.

Nature·2026
Same author

ATP is dispensable for E. coli DNA replication and eukaryotic helicase activity.

Nature communications·2026
Same author

Redesign of energetically frustrated regions rescues function in defective T4 clamp loaders.

bioRxiv : the preprint server for biology·2026
Same author

The E. coli DnaX clamp loader sharply bends DNA to load β-clamp at nicks and small gaps.

Molecular cell·2026

DNA polymerases stall at G-quadruplex structures, which are obstacles to genome duplication. This study reveals polymerases repeatedly bind and unbind when encountering these DNA roadblocks.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Faithful genome duplication relies on DNA replication proteins overcoming obstacles like G-quadruplexes.
  • G-quadruplexes are higher-order DNA structures that impede genomic maintenance.
  • Understanding protein-DNA interactions at these structures is crucial.

Purpose of the Study:

  • To develop a real-time method for observing DNA polymerase interactions with G-quadruplexes.
  • To investigate how DNA polymerases behave when encountering G-quadruplex obstacles.
  • To provide a versatile single-molecule assay for studying DNA-protein-obstacle interactions.

Main Methods:

  • Real-time fluorescence microscopy in a microfluidic flow cell.
  • Immobilization of primed DNA oligonucleotides containing G-quadruplexes on functionalized glass coverslips.

More Related Videos

Author Spotlight: Unraveling the Dynamics of Eukaryotic DNA Replication Through Single-Molecule Visualization
07:37

Author Spotlight: Unraveling the Dynamics of Eukaryotic DNA Replication Through Single-Molecule Visualization

Published on: September 27, 2024

1.3K
Single-molecule Manipulation of G-quadruplexes by Magnetic Tweezers
08:28

Single-molecule Manipulation of G-quadruplexes by Magnetic Tweezers

Published on: September 19, 2017

7.9K

Related Experiment Videos

Last Updated: May 10, 2025

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

267
Author Spotlight: Unraveling the Dynamics of Eukaryotic DNA Replication Through Single-Molecule Visualization
07:37

Author Spotlight: Unraveling the Dynamics of Eukaryotic DNA Replication Through Single-Molecule Visualization

Published on: September 27, 2024

1.3K
Single-molecule Manipulation of G-quadruplexes by Magnetic Tweezers
08:28

Single-molecule Manipulation of G-quadruplexes by Magnetic Tweezers

Published on: September 19, 2017

7.9K
  • Monitoring of fluorescently labeled DNA polymerases over time.

    • Main Results:

    • Successfully visualized DNA polymerase stalling at G-quadruplex structures.
    • Observed that yeast polymerase δ exhibits a continuous binding and unbinding cycle when encountering a G-quadruplex.
    • Demonstrated the utility of the single-molecule assay for studying polymerase dynamics.

    Conclusions:

    • The developed method allows real-time observation of DNA polymerase behavior at G-quadruplexes.
    • DNA polymerases engage in dynamic binding-unbinding cycles when stalled by G-quadruplexes.
    • This assay platform can be adapted to study various DNA-maintenance proteins and DNA obstacles.