Related Experiment Video

Updated: May 28, 2026

Two- and Three-Dimensional Live Cell Imaging of DNA Damage Response Proteins
10:24

Two- and Three-Dimensional Live Cell Imaging of DNA Damage Response Proteins

Published on: September 28, 2012

A genome-wide screen identifies p97 as an essential regulator of DNA damage-dependent CDT1 destruction

Malavika Raman1, Courtney G Havens, Johannes C Walter

  • 1Department of Cell Biology, Harvard Medical School, Boston, MA 01230, USA.

Molecular Cell
|October 11, 2011
PubMed

Insights

The CRL4(CDT2) E3 ligase degrades CDT1 and SET8 proteins during DNA repair. Nucleotide excision repair factors and the p97 AAA(+)-ATPase complex are crucial for this process, ensuring timely protein removal from chromatin.

Area of Science:

  • Cellular Biology
  • Molecular Biology
  • Biochemistry

Background:

  • The E3 ubiquitin ligase CRL4(CDT2) targets proteins like CDT1 and SET8 for degradation during DNA replication and repair.
  • CRL4(CDT2) activity is linked to DNA repair through its interaction with chromatin-bound PCNA.

Purpose of the Study:

  • To identify factors involved in CDT1 destruction after UV irradiation using a genome-wide siRNA screen.
  • To elucidate the mechanisms regulating CRL4(CDT2) substrate degradation.

Main Methods:

  • Genome-wide siRNA screen to identify factors affecting CDT1 destruction.
  • Investigation of the roles of nucleotide excision repair factors, COP9/Signalosome, and p97 AAA(+)-ATPase complex.
  • In vivo and in vitro (Xenopus egg extract) degradation assays.

Main Results:

  • Multiple factors, including nucleotide excision repair factors, are required for UV-induced CDT1 destruction.
  • Nucleotide excision repair factors recruit PCNA to damaged DNA, promoting CDT1 degradation.
  • COP9/Signalosome regulates CDT2 stability via CUL4 deneddylation.
  • The p97 AAA(+)-ATPase/UFD1 complex mediates the removal and degradation of ubiquitinated CDT1 and SET8 from chromatin.

Conclusions:

  • This study identifies key regulators of CRL4(CDT2) substrate degradation during DNA repair.
  • The findings provide a comprehensive resource for understanding the pathways controlling chromatin-associated protein turnover.

More Related Videos

Detection and Visualization of DNA Damage-induced Protein Complexes in Suspension Cell Cultures Using the Proximity Ligation Assay13:10

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

Published on: June 9, 2017

Quantitative Detection of DNA-Protein Crosslinks and Their Post-Translational Modifications10:12

Quantitative Detection of DNA-Protein Crosslinks and Their Post-Translational Modifications

Published on: April 21, 2023

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...
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.
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...
Abnormal Proliferation02:23

Abnormal Proliferation

Under normal conditions, most adult cells remain in a non-proliferative state unless stimulated by internal or external factors to replace lost cells. Abnormal cell proliferation is a condition in which the cell's growth exceeds and is uncoordinated with normal cells. In such situations, cell division persists in the same excessive manner even after cessation of the stimuli, leading to persistent tumors. The tumor arises from the damaged cells that replicate to pass the damage to the daughter...
Nucleotide Excision Repair01:38

Nucleotide Excision Repair

DNA Distortion and Damage
Cells are regularly exposed to mutagens—factors in the environment that can damage DNA and generate mutations. UV radiation is one of the most common mutagens and is estimated to introduce a significant number of changes in DNA. These include bends or kinks in the structure, which can block DNA replication or transcription. If these errors are not fixed, the damage can cause mutations, which in turn can result in cancer or disease depending on which sequences are...