Related Experiment Video

Updated: Aug 10, 2026

Detection and Visualization of DNA Damage-induced Protein Complexes in Suspension Cell Cultures Using the Proximity Ligation Assay
13:10

Detection and Visualization of DNA Damage-induced Protein Complexes in Suspension Cell Cultures Using the Proximity Ligation Assay

Published on: June 9, 2017

Selective induction of E2F1 in response to DNA damage, mediated by ATM-dependent phosphorylation

W C Lin1, F T Lin, J R Nevins

  • 1Department of Genetics, Howard Hughes Medical Institute, Durham, North Carolina 27710, USA.

Genes & Development
|July 19, 2001
PubMed

Insights

DNA damage triggers the accumulation of E2F1, a protein crucial for apoptosis. This process relies on ATM kinase activity and specific phosphorylation, highlighting the Rb/E2F pathway

Area of Science:

  • Molecular Biology
  • Cell Cycle Regulation
  • DNA Damage Response

Background:

  • The p53 protein mediates apoptosis following DNA damage and Rb cell cycle pathway deregulation.
  • The Rb-regulated E2F1 protein is implicated in inducing apoptosis and p53 accumulation.

Purpose of the Study:

  • To investigate the role of E2F1 in the cellular response to DNA damage.
  • To elucidate the mechanisms regulating E2F1 accumulation and its function in apoptosis.

Main Methods:

  • Analysis of E2F1 accumulation in response to DNA damage.
  • Investigation of ATM and ATR kinase activity in E2F1 phosphorylation and stabilization.
  • Site-directed mutagenesis to identify critical phosphorylation sites on E2F1.
  • Assessment of E2F1's requirement for apoptosis in mouse thymocytes.

Main Results:

  • DNA damage specifically induces E2F1 accumulation, dependent on ATM kinase activity.
  • ATM and ATR kinases phosphorylate E2F1 at a specific N-terminal site, essential for its ATM-mediated stabilization.
  • E2F1 is indispensable for DNA damage-induced apoptosis in mouse thymocytes.

Conclusions:

  • The cellular DNA damage response utilizes signals from the Rb/E2F cell cycle pathway.
  • E2F1 acts as a key mediator linking DNA damage, cell cycle control, and apoptosis.

More Related Videos

Visualization of DNA Repair Proteins Interaction by Immunofluorescence07:55

Visualization of DNA Repair Proteins Interaction by Immunofluorescence

Published on: June 26, 2020

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

Negative Regulator Molecules01:23

Negative Regulator Molecules

Positive regulators allow a cell to advance through cell cycle checkpoints. Negative regulators have an equally important role as they terminate a cell’s progression through the cell cycle—or pause it—until the cell meets specific criteria.
Long-patch Base Excision Repair01:02

Long-patch Base Excision Repair

Since the discovery of the two BER pathways, there has been a debate about how a cell chooses one pathway over the other and the factors determining this selection. Numerous in vitro experiments have pointed out multiple determinants for the sub-pathway selection. These are:
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...
Inhibition of Cdk Activity02:34

Inhibition of Cdk Activity

The orderly progression of the cell cycle depends on the activation of Cdk protein by binding to its cyclin partner. However, the cell cycle must be restricted when undergoing abnormal changes. Most cancers correlate to the deregulated cell cycle, and since Cdks are a central component of the cell cycle, Cdk inhibitors are extensively studied to develop anticancer agents. For instance, cyclin D associates with several Cdks, such as Cdk 4/6, to form an active complex. The cyclin D-Cdk4/6 complex...
Mitogens and the Cell Cycle02:38

Mitogens and the Cell Cycle

Mitogens and their receptors play a crucial role in controlling the progression of the cell cycle. However, the loss of mitogenic control over cell division leads to tumor formation. Therefore, mitogens and mitogen receptors play an important role in cancer research. For instance, the epidermal growth factor (EGF) - a type of mitogen and its transmembrane receptor (EGFR), decides the fate of the cell's proliferation. When EGF binds to EGFR, a member of the ErbB family of tyrosine kinase...
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...