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 Concept Videos

Replication in Eukaryotes02:31

Replication in Eukaryotes

Overview
Replication in Eukaryotes01:29

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...
Replication in Eukaryotes02:31

Replication in Eukaryotes

Overview
Replication in Eukaryotes01:29

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...
DNA Replication02:40

DNA Replication

DNA replication involves the separation of the two strands of the double helix, with each strand serving as a template from which the new complementary strand is copied.  After replication, each double-stranded DNA includes one parental or “old” strand and one “new” strand. This is known as semiconservative replication. The resulting DNA molecules have the same sequence and are divided equally into the two daughter cells.
Replication in Prokaryotes
DNA replication uses a large number of...
Lagging Strand Synthesis01:59

Lagging Strand Synthesis

During replication, the complementary strands in double-stranded DNA are synthesized at different rates. Replication first begins on the leading strand. Replication starts later, occurs more slowly, and proceeds discontinuously on the lagging strand.
There are several major differences between synthesis of the leading strand and synthesis of the lagging strand. 1) Leading strand synthesis happens in the direction of replication fork opening, whereas lagging strand synthesis happens in the...

You might also read

Related Articles

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

Sort by
Same author

Biochemical impact of p300-mediated acetylation of replication protein A: Implications for DNA metabolic pathway choice.

The Journal of biological chemistry·2025
Same author

Simian virus 40 (SV40) - Fresh perspectives on a historic virus.

Virology·2025
Same author

Analyzing DNA-Protein Interactions with Streptavidin-Based Biolayer Interferometry.

Journal of visualized experiments : JoVE·2025
Same author

The effect of replication protein A inhibition and post-translational modification on ATR kinase signaling.

Scientific reports·2024
Same author

The Effect of Replication Protein A Inhibition and Post-Translational Modification on ATR Kinase Signaling.

Research square·2024
Same author

Human CST Stimulates Base Excision Repair to Prevent the Accumulation of Oxidative DNA Damage.

Journal of molecular biology·2024

Related Experiment Video

Updated: Jun 15, 2026

Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase
07:27

Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase

Published on: April 29, 2010

Reconstitution of eukaryotic lagging strand DNA replication.

Lata Balakrishnan1, Jason W Gloor, Robert A Bambara

  • 1Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, NY 14642, USA.

Methods (San Diego, Calif.)
|February 25, 2010
PubMed
Summary

Okazaki fragment maturation in eukaryotic DNA replication relies on Flap endonuclease 1 (FEN1) and other proteins. This review details in vitro methods to study these complex DNA replication processes.

More Related Videos

Kinetics of Lagging-strand DNA Synthesis In Vitro by the Bacteriophage T7 Replication Proteins
08:14

Kinetics of Lagging-strand DNA Synthesis In Vitro by the Bacteriophage T7 Replication Proteins

Published on: February 25, 2017

Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
08:53

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

Published on: May 2, 2025

Related Experiment Videos

Last Updated: Jun 15, 2026

Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase
07:27

Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase

Published on: April 29, 2010

Kinetics of Lagging-strand DNA Synthesis In Vitro by the Bacteriophage T7 Replication Proteins
08:14

Kinetics of Lagging-strand DNA Synthesis In Vitro by the Bacteriophage T7 Replication Proteins

Published on: February 25, 2017

Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
08:53

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

Published on: May 2, 2025

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Eukaryotic DNA replication ensures genome integrity through precise DNA synthesis.
  • Lagging strand synthesis involves Okazaki fragment processing, a complex pathway.
  • Flap endonuclease 1 (FEN1) is crucial for Okazaki fragment maturation.

Purpose of the Study:

  • To review the enzymatic steps in Okazaki fragment maturation.
  • To describe methods for studying DNA replication intermediates in vitro.
  • To present protocols for reconstituting lagging strand replication.

Main Methods:

  • Design of specific DNA substrates simulating maturation intermediates.
  • In vitro reconstitution reactions for partial and complete lagging strand replication.
  • Utilizing genetic studies and enzymatic assays to understand protein functions.

Main Results:

  • FEN1 cleaves flaps displaced by DNA polymerase delta (pol delta).
  • Pif1 and Dna2 helicases/nucleases elongate flaps, contributing to maturation.
  • In vitro reconstitution reactions have elucidated mechanistic models of lagging strand synthesis.

Conclusions:

  • Okazaki fragment maturation is a complex process involving multiple proteins.
  • FEN1, Pif1, and Dna2 play vital roles in lagging strand DNA replication.
  • In vitro reconstitution provides key insights into DNA replication mechanisms.