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

Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

5.9K
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,...
5.9K
DNA Damage can Stall the Cell Cycle02:37

DNA Damage can Stall the Cell Cycle

9.3K
In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
9.3K
The DNA Replication Fork01:02

The DNA Replication Fork

36.7K
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...
36.7K
The Spindle Assembly Checkpoint02:19

The Spindle Assembly Checkpoint

3.2K
The spindle assembly checkpoint is a molecular surveillance mechanism ensuring the fidelity of chromosome segregation during anaphase. The checkpoint monitors the completion of all the prerequisite steps before chromosome segregation to determine whether the segregation process should proceed or be delayed.
Many proteins function together to control the spindle assembly checkpoint. Mutations affecting these proteins may allow cells to proceed into anaphase prematurely, resulting in the...
3.2K
Homologous Recombination02:31

Homologous Recombination

51.4K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
51.4K
S-Cdk Initiates DNA Replication02:38

S-Cdk Initiates DNA Replication

4.8K
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...
4.8K

You might also read

Related Articles

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

Sort by
Same author

Understanding single stranded DNA gaps: from formation to fate.

The Biochemical journal·2026
Same author

Genome-wide CRISPR screens identify the EXO1-CAF-1 pathway suppressing R-loop-associated DNA damage.

Nucleic acids research·2026
Same author

The nuclease EXO1 promotes genomic instability by degrading nascent DNA in BRCA-proficient cells.

Nature communications·2026
Same author

The RRM domains of PARP14 mediate replication fork degradation in BRCA2-deficient cells.

NAR cancer·2026
Same author

Translesion-synthesis-mediated bypass of DNA lesions occurs predominantly behind replication forks restarted by PrimPol.

Cell reports·2025
Same author

CAF-1 promotes efficient PrimPol recruitment to nascent DNA for single-stranded DNA gap formation.

Nucleic acids research·2024

Related Experiment Video

Updated: Sep 7, 2025

Author Spotlight: Unveiling the Role of SNF2L in Replication Fork Stability and Genome Duplication
05:55

Author Spotlight: Unveiling the Role of SNF2L in Replication Fork Stability and Genome Duplication

Published on: August 23, 2024

638

The TIP60-ATM axis regulates replication fork stability in BRCA-deficient cells.

Emily M Schleicher1, Ashna Dhoonmoon1, Lindsey M Jackson1

  • 1Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, PA, 17033, USA.

Oncogenesis
|June 18, 2022
PubMed
Summary

The TIP60-ATM pathway drives replication fork reversal and degradation in BRCA-deficient cells. Genetic screens reveal DNA repair genes critical for survival during ATM inhibition, especially in BRCA2-deficient contexts.

More Related Videos

Detection of Post-Replicative Gaps Accumulation and Repair in Human Cells Using the DNA Fiber Assay
10:32

Detection of Post-Replicative Gaps Accumulation and Repair in Human Cells Using the DNA Fiber Assay

Published on: February 3, 2022

6.6K
Characterizing DNA Repair Processes at Transient and Long-lasting Double-strand DNA Breaks by Immunofluorescence Microscopy
08:31

Characterizing DNA Repair Processes at Transient and Long-lasting Double-strand DNA Breaks by Immunofluorescence Microscopy

Published on: June 8, 2018

9.2K

Related Experiment Videos

Last Updated: Sep 7, 2025

Author Spotlight: Unveiling the Role of SNF2L in Replication Fork Stability and Genome Duplication
05:55

Author Spotlight: Unveiling the Role of SNF2L in Replication Fork Stability and Genome Duplication

Published on: August 23, 2024

638
Detection of Post-Replicative Gaps Accumulation and Repair in Human Cells Using the DNA Fiber Assay
10:32

Detection of Post-Replicative Gaps Accumulation and Repair in Human Cells Using the DNA Fiber Assay

Published on: February 3, 2022

6.6K
Characterizing DNA Repair Processes at Transient and Long-lasting Double-strand DNA Breaks by Immunofluorescence Microscopy
08:31

Characterizing DNA Repair Processes at Transient and Long-lasting Double-strand DNA Breaks by Immunofluorescence Microscopy

Published on: June 8, 2018

9.2K

Area of Science:

  • Molecular Biology
  • Genetics
  • DNA Repair

Background:

  • Replication fork stability is crucial for genome integrity.
  • Stalled forks can be reversed by translocases like SMARCAL1 and degraded if not protected by the BRCA pathway.
  • ATM and ATR kinases are key regulators of DNA damage response.

Purpose of the Study:

  • To investigate the role of the TIP60-ATM pathway in replication fork reversal.
  • To identify genetic factors influencing cellular sensitivity to ATM inhibitors in wild-type and BRCA2-knockout cells.

Main Methods:

  • Investigated the TIP60-ATM pathway's role in recruiting SMARCAL1 to stalled forks.
  • Conducted genome-wide CRISPR knockout screens to assess sensitivity to ATM inhibitors.
  • Validated top genetic hits from screens in both wild-type and BRCA2-knockout cells.

Main Results:

  • The TIP60-ATM pathway promotes replication fork reversal by recruiting SMARCAL1, leading to degradation in BRCA-deficient cells.
  • ATM activity in fork reversal is distinct from ATR.
  • Identified common genes regulating responses to multiple ATM inhibitors.
  • Discovered that DNA repair genes (RAD17, MDC1, USP28) are essential for survival upon ATM inhibition in BRCA2-knockout cells, unlike in wild-type cells.

Conclusions:

  • The TIP60-ATM pathway plays a specific role in replication fork reversal and degradation, particularly in BRCA-deficient cells.
  • ATM inhibitors exhibit differential efficacy based on BRCA status, with DNA repair pathways being critical in BRCA2-deficient contexts.
  • Findings may inform the clinical use of ATM inhibitors, especially for BRCA-mutated cancers.