Related Experiment Video

Updated: May 7, 2026

Visualization of DNA Repair Proteins Interaction by Immunofluorescence
07:55

Visualization of DNA Repair Proteins Interaction by Immunofluorescence

Published on: June 26, 2020

Damage-induced DNA replication stalling relies on MAPK-activated protein kinase 2 activity

Frederik Köpper1, Cathrin Bierwirth, Margarete Schön

  • 1Institute of Molecular Oncology and Göttingen Centre of Molecular Biosciences, Faculty of Medicine, University of Göttingen, 37077 Göttingen, Germany.

Proceedings of the National Academy of Sciences of the United States of America
|October 2, 2013
PubMed

Insights

MAP kinase-activated protein kinase 2 (MK2) plays a key role in DNA damage response. MK2 inhibition protects cells from DNA damage and rescues replication forks, revealing its importance in cell survival signaling.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • DNA damage can stall replication forks, leading to replication stress.
  • Kinases are crucial in managing cellular responses to DNA damage and replication stress.

Purpose of the Study:

  • To identify kinases involved in the accumulation of phosphohistone 2AX (γH2AX) during UV-induced replication stress.
  • To investigate the role of MAP kinase-activated protein kinase 2 (MK2) in DNA damage response and replication fork stability.

Main Methods:

  • siRNA screening to identify key kinases.
  • Analysis of phosphohistone 2AX (γH2AX) accumulation.
  • Cell viability assays and apoptosis measurements in MK2-deficient mice.
  • DNA fiber assays to assess replication fork dynamics.
  • Treatment with gemcitabine and Chk1 antagonism.

Main Results:

  • Knockdown of MK2 significantly reduced γH2AX accumulation after UV irradiation.
  • MK2 depletion or inhibition conferred resistance to DNA damage-induced cell death.
  • MK2 deficiency reduced UV-induced apoptosis in mouse skin.
  • MK2 activity was essential for DNA damage response, ssDNA accumulation, and reduced cell survival upon gemcitabine treatment or Chk1 inhibition.
  • MK2 inhibition or knockdown rescued impaired DNA replication, dependent on translesion DNA polymerases.

Conclusions:

  • MK2 signaling is a critical regulator of replication fork dynamics and origin firing following DNA damage.
  • MK2 inhibition offers a potential therapeutic strategy to protect against DNA damage and enhance cancer treatment efficacy.

More Related Videos

Application of Stopped-flow Kinetics Methods to Investigate the Mechanism of Action of a DNA Repair Protein11:01

Application of Stopped-flow Kinetics Methods to Investigate the Mechanism of Action of a DNA Repair Protein

Published on: March 31, 2010

Visualizing and Quantifying Endonuclease-Based Site-Specific DNA Damage10:59

Visualizing and Quantifying Endonuclease-Based Site-Specific DNA Damage

Published on: August 21, 2021

Related Concept Videos

DNA Damage can Stall the Cell Cycle02:36

DNA Damage can Stall the Cell Cycle

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...
DNA Damage Can Stall the Cell Cycle02:36

DNA Damage Can Stall the Cell Cycle

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...
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

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, a...
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

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, a...
The DNA Replication Fork01:02

The DNA Replication Fork

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 forks, one in...
S-Cdk Initiates DNA Replication02:38

S-Cdk Initiates DNA Replication

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 replication.