Related Experiment Video

Updated: Jan 15, 2026

Author Spotlight: Quantitative Detection of DNA Protein Crosslinks and Their Post-Translational Modifications
10:12

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

Published on: April 21, 2023

3.6K

CRL4 mediates autoubiquitination of DDB1 upon deneddylation inhibition

Yeong-Mu Kim1, Jae-Hyun Jo1, Dong-Kyu Kim1

  • 1Department of Biochemistry, Chungbuk National University, Cheongju, 28644, South Korea; Department of Biological Sciences and Biotechnology, Chungbuk National University, Cheongju, 28644, South Korea.

Biochemical and Biophysical Research Communications
|October 9, 2025
PubMed

Insights

Inhibiting deneddylation destabilizes DDB1 via CRL4-dependent autoubiquitination, impairing DNA replication and enhancing vulnerability to p97/VCP inhibitors. This reveals a synthetic interaction between CRL4 and p97/VCP.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Cullin-RING E3 ubiquitin ligases (CRLs) regulate protein homeostasis through neddylation.
  • The CRL4 complex, featuring DDB1 (damage-specific DNA binding protein 1), is crucial for DNA replication, proliferation, and repair.
  • DDB1 acts as an adaptor, linking CUL4A/B to substrate receptors (DCAFs).

Purpose of the Study:

  • To investigate the impact of deneddylation inhibition on CRL4 complex stability and function.
  • To elucidate the mechanism by which deneddylation inhibition affects DDB1 levels.
  • To explore the functional consequences of DDB1 loss and its interaction with other cellular pathways.

Main Methods:

  • Inhibition of deneddylation using specific inhibitors (CSN5i).
  • Assessment of DDB1 stability and CRL4 autoubiquitination.
  • Analysis of CRL4 recruitment to chromatin.
  • Phenotypic analysis of cells under deneddylation inhibition.
  • Evaluation of cellular vulnerability to p97/VCP inhibitors.

Main Results:

  • Inhibition of deneddylation leads to destabilization of DDB1 through CRL4-dependent autoubiquitination.
  • DDB1 loss compromises CRL4 chromatin recruitment, impairing DNA replication.
  • A cellular phenotype resembling RepID deficiency arises despite normal RepID expression.
  • Deneddylation inhibition sensitizes cells to p97/VCP inhibition, indicating a synthetic interaction.

Conclusions:

  • Deneddylation inhibition triggers CRL4-mediated DDB1 autoubiquitination and degradation.
  • Disruption of CRL4 homeostasis impacts DNA replication and cellular function.
  • A novel synthetic lethality is identified between CRL4 pathway disruption and p97/VCP inhibition.
Keywords:
CRL4 autoubiquitinationCSN5 inhibitorDDB1RepIDp97/VCP inhibitor

More Related Videos

In Vitro Ubiquitination and Deubiquitination Assays of Nucleosomal Histones11:36

In Vitro Ubiquitination and Deubiquitination Assays of Nucleosomal Histones

Published on: July 25, 2019

11.4K
Chemical Inactivation of the E3 Ubiquitin Ligase Cereblon by Pomalidomide-based Homo-PROTACs10:44

Chemical Inactivation of the E3 Ubiquitin Ligase Cereblon by Pomalidomide-based Homo-PROTACs

Published on: May 15, 2019

13.7K

Related Concept Videos

Anaphase Promoting Complex00:50

Anaphase Promoting Complex

The stepwise destruction of specific proteins is necessary for the progression and completion of the cell cycle. Such proteins are ubiquitinated by ubiquitin ligases and then subsequently destroyed by the proteasome. The SCF (Skp1/Cullin/F-box) and the anaphase-promoting complex (APC) are two important ubiquitin ligases involved in cell cycle progression. While SCF is active throughout the cell cycle, APC gets activated during metaphase to anaphase transition. Cdc20 or Cdh1 binds to APC and...
3.3K
Regulated Protein Degradation02:58

Regulated Protein Degradation

It is vital to regulate the activity of enzymatic as well as non-enzymatic proteins inside the cell. This can be achieved either through creating a balance between their rate of synthesis and degradation or regulating the intrinsic activity of the protein. Both these regulation mechanisms play an essential role in the normal functioning of cells.
Protein degradation plays two important roles in the cells. It helps to protect cells from misfolded or damaged proteins before they lead to a...
8.7K
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...
10.0K
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...
3.0K
Covalently Linked Protein Regulators02:04

Covalently Linked Protein Regulators

Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein....
8.7K
Receptor Downregulation in MVBs01:15

Receptor Downregulation in MVBs

Multivesicular bodies (MVBs) are mature endosomes that sort ubiquitinated proteins and then fuse with lysosomes to degrade the sorted proteins. Epidermal growth factor (EGF) and its receptor (EGFR) form a complex that can be internalized through endocytosis, sorted into an MVB, and later degraded.
The EGFR can initiate signaling pathways that  lead to cell proliferation, migration, and differentiation. Overexpression of EGFR  stimulates cells to proliferate. Excessive  EGFR...
2.8K