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

The DNA Replication Fork01:02

The DNA Replication Fork

36.1K
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...
36.1K
S-Cdk Initiates DNA Replication02:38

S-Cdk Initiates DNA Replication

4.7K
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...
4.7K
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

5.8K
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,...
5.8K
Replication in Prokaryotes01:32

Replication in Prokaryotes

25.0K
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...
25.0K
Translesion DNA Polymerases02:10

Translesion DNA Polymerases

10.0K
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...
10.0K
The Replisome03:01

The Replisome

33.7K
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...
33.7K

You might also read

Related Articles

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

Sort by
Same author

Cohesin residence time gates 3D genome response to histone hyperacetylation.

bioRxiv : the preprint server for biology·2026
Same author

A CDK1 phospho-switch reprograms TRAIP to unload replisomes in mitosis.

Science (New York, N.Y.)·2026
Same author

A progeria syndrome links DNA hypermethylation to age-related pathology.

Nature genetics·2026
Same author

Distinct roles of MCM2-7 subunits in replication licensing in human cells.

Nature communications·2026
Same author

In silico discovery of nanobody binders to a G-protein coupled receptor using AlphaFold-Multimer.

Nature communications·2026
Same author

Cohesin prevents local mixing of condensed euchromatic domains in living human cells.

bioRxiv : the preprint server for biology·2026

Related Experiment Video

Updated: Jul 19, 2025

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

411

In silico protein interaction screening uncovers DONSON's role in replication initiation.

Yang Lim1, Lukas Tamayo-Orrego2, Ernst Schmid1

  • 1Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Blavatnik Institute, Boston, MA 02115, USA.

Science (New York, N.Y.)
|August 17, 2023
PubMed
Summary
This summary is machine-generated.

DONSON protein is crucial for eukaryotic DNA replication initiation by assembling the CDC45-MCM2-7-GINS (CMG) helicase. Mutations in DONSON cause microcephalic dwarfism, highlighting its role in human development.

More Related Videos

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique
07:18

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

Published on: October 27, 2011

40.0K
Pulldown Assay Coupled with Co-Expression in Bacteria Cells as a Time-Efficient Tool for Testing Challenging Protein-Protein Interactions
07:03

Pulldown Assay Coupled with Co-Expression in Bacteria Cells as a Time-Efficient Tool for Testing Challenging Protein-Protein Interactions

Published on: December 23, 2022

3.0K

Related Experiment Videos

Last Updated: Jul 19, 2025

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

411
Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique
07:18

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

Published on: October 27, 2011

40.0K
Pulldown Assay Coupled with Co-Expression in Bacteria Cells as a Time-Efficient Tool for Testing Challenging Protein-Protein Interactions
07:03

Pulldown Assay Coupled with Co-Expression in Bacteria Cells as a Time-Efficient Tool for Testing Challenging Protein-Protein Interactions

Published on: December 23, 2022

3.0K

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Eukaryotic DNA replication initiation relies on the assembly of the CDC45-MCM2-7-GINS (CMG) helicase.
  • In yeast, a pre-loading complex (pre-LC) facilitates GINS binding to chromatin-associated MCM2-7 to form CMG.
  • The precise mechanism of CMG assembly in vertebrates remains incompletely understood.

Purpose of the Study:

  • To investigate the role of the metazoan protein DONSON in CMG helicase assembly in vertebrates.
  • To elucidate the function of DONSON in the context of replication initiation and its potential link to microcephalic primordial dwarfism.

Main Methods:

  • Utilized AlphaFold for in silico screening of protein-protein interactions.
  • Performed experimental validation of predicted protein interactions.
  • Investigated the impact of a patient-derived DONSON mutation on CMG assembly and its phenotypic consequences in a mouse model.

Main Results:

  • DONSON acts as a scaffold for a vertebrate pre-LC, including GINS, TOPBP1, and DNA polymerase epsilon.
  • DONSON facilitates the docking of the pre-LC onto MCM2-7, thereby delivering GINS for CMG formation.
  • A patient-derived DONSON mutation impairs CMG assembly and causes microcephalic dwarfism in mice.

Conclusions:

  • DONSON is essential for CMG helicase assembly in vertebrates, unifying understanding of eukaryotic replication initiation.
  • Defective CMG assembly due to DONSON mutations is implicated in microcephalic dwarfism.
  • In silico protein-protein interaction screening, exemplified by AlphaFold, accelerates mechanistic biological discovery.