Related Experiment Video

Updated: Jun 13, 2026

A High Resolution Method to Monitor Phosphorylation-dependent Activation of IRF3
11:44

A High Resolution Method to Monitor Phosphorylation-dependent Activation of IRF3

Published on: January 24, 2016

The Cullin 3 substrate adaptor KLHL20 mediates DAPK ubiquitination to control interferon responses

Yu-Ru Lee1, Wei-Chien Yuan, Hsuan-Chung Ho

  • 1Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan.

The EMBO Journal
|April 15, 2010
PubMed

Insights

The BTB-Kelch protein KLHL20 targets death-associated protein kinase (DAPK) for degradation. Interferon (IFN) blocks this, stabilizing DAPK and enhancing cancer cell death, potentially overcoming IFN resistance.

Area of Science:

  • Molecular Biology
  • Cellular Biology
  • Cancer Research

Background:

  • Death-associated protein kinase (DAPK) mediates interferon (IFN)-induced cell death.
  • The precise mechanisms by which IFN regulates DAPK activation are not fully understood.
  • Understanding DAPK regulation is crucial for cancer therapy involving IFNs.

Purpose of the Study:

  • To identify novel regulators of DAPK.
  • To elucidate the mechanism of DAPK regulation by interferons.
  • To explore the role of DAPK stabilization in IFN responsiveness and cancer therapy.

Main Methods:

  • Identification of KLHL20 as a DAPK-binding protein.
  • Characterization of the KLHL20-Cullin 3 (Cul3)-ROC1 E3 ligase complex.
  • Analysis of DAPK ubiquitination, degradation, and stabilization in response to IFN.

Main Results:

  • KLHL20 targets DAPK for polyubiquitination and proteasomal degradation via the KLHL20-Cul3-ROC1 E3 ligase complex.
  • IFN treatment causes KLHL20 to sequester in PML nuclear bodies, separating it from DAPK.
  • IFN-induced sequestration of KLHL20 leads to DAPK stabilization, enhancing IFN responsiveness and promoting cell death.

Conclusions:

  • KLHL20 acts as a negative regulator of DAPK by promoting its degradation.
  • IFN-induced sequestration of KLHL20 is a key mechanism for DAPK stabilization.
  • This pathway is critical for IFN-induced apoptosis and autophagy, offering potential strategies to overcome IFN resistance in cancer.

More Related Videos

Using In Vitro Fluorescence Resonance Energy Transfer to Study the Dynamics Of Protein Complexes at a Millisecond Time Scale10:50

Using In Vitro Fluorescence Resonance Energy Transfer to Study the Dynamics Of Protein Complexes at a Millisecond Time Scale

Published on: March 14, 2019

In vivo and in vitro Studies of Adaptor-clathrin Interaction17:14

In vivo and in vitro Studies of Adaptor-clathrin Interaction

Published on: January 26, 2011

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...
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 activation may...
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...
Regulation of the Unfolded Protein Response01:31

Regulation of the Unfolded Protein Response

Inositol-requiring kinase one or IRE1 is the most conserved eukaryotic unfolded protein response (UPR) receptor. It is a type I transmembrane protein kinase receptor with a distinctive site-specific RNase activity. As the binding mechanics of the misfolded proteins with the N-terminal domain of IRE-1 are unclear, three binding models — direct, indirect, and allosteric -- are proposed for receptor activation. Nevertheless, it is known that once a misfolded protein associates with IRE1, it...
The Unfolded Protein Response01:37

The Unfolded Protein Response

The ER is the hub of protein synthesis in a cell. It has robust systems to quality control protein folding and also for degradation of terminally misfolded proteins. Under normal conditions, a small proportion of misfolded proteins that cannot be salvaged need to be transported to the cytoplasm by the ER-associated degradation or ERAD pathways. However, if the ERAD cannot handle the misfolded proteins, the cell activates the unfolded protein response or UPR to adjust the protein folding...
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...