Related Experiment Video

Updated: Jun 1, 2026

Production, Crystallization, and Structure Determination of the IKK-binding Domain of NEMO
13:02

Production, Crystallization, and Structure Determination of the IKK-binding Domain of NEMO

Published on: December 28, 2019

NF-κB essential modulator (NEMO) interaction with linear and lys-63 ubiquitin chains contributes to NF-κB activation

Kamyar Hadian1, Richard A Griesbach, Scarlett Dornauer

  • 1Department of Cellular Signal Integration Helmholtz Zentrum München, German Research Center for Environmental Health, Institute of Toxicology, Neuherberg, Germany. daniel.krappmann@helmholtz-muenchen.de

The Journal of Biological Chemistry
|May 31, 2011
PubMed

Insights

The NEMO protein binds to both linear and Lys-63 ubiquitin chains, which is crucial for activating the IKK complex and NF-κB signaling pathways. Both chain types cooperate for optimal IKK activation.

Area of Science:

  • Molecular Biology
  • Cell Signaling
  • Immunology

Background:

  • The IκB kinase (IKK) complex regulates canonical NF-κB signaling.
  • Activation of IKK requires the IKK regulatory subunit IKKγ/NEMO to sense ubiquitin chains.

Purpose of the Study:

  • To investigate whether NEMO binding to Lys-63-linked or linear ubiquitin chains is critical for IKK activation.
  • To elucidate the role of NEMO's interaction with different ubiquitin chain types in NF-κB signaling.

Main Methods:

  • Assessed NEMO's binding preference for linear versus Lys-63 ubiquitin chains.
  • Utilized site-directed mutagenesis in NEMO to selectively disrupt binding to linear ubiquitin chains.
  • Analyzed NF-κB signaling, target gene expression, and apoptosis induction in response to TNFα stimulation in fibroblasts and T cells.

Main Results:

  • NEMO's C terminus preferentially binds linear ubiquitin chains.
  • Immobilization of NEMO enhances its interaction with Lys-63 ubiquitin chains.
  • Mutations impairing linear ubiquitin binding partially compromised TNFα-induced NF-κB signaling and apoptosis.

Conclusions:

  • NEMO's interaction with both linear and Lys-63 ubiquitin chains is essential for optimal IKK activation.
  • Both ubiquitin chain types cooperate to trigger canonical NF-κB signaling in vivo.

More Related Videos

NF-κB-dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells10:57

NF-κB-dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells

Published on: January 12, 2020

A Guide to Production, Crystallization, and Structure Determination of Human IKK1/α11:27

A Guide to Production, Crystallization, and Structure Determination of Human IKK1/α

Published on: November 2, 2018

Related Concept Videos

NF-κB-dependent Signaling Pathway02:26

NF-κB-dependent Signaling Pathway

The transcription factor NF-κB was discovered in 1986 in the lab of Nobel laureate Professor David Baltimore, for its interaction with the immunoglobulin light chain enhancer in B-cells. After more than three decades of study, it is now evident that NF-κB regulates the expression of over 100 genes. Most of these genes play an essential role in the innate and adaptive immune responses as well as the inflammatory responses of animals.
NF-κB-dependent Signaling Mechanism
The heterodimer of NF-κB...
Regulation of Nuclear Protein Sorting01:45

Regulation of Nuclear Protein Sorting

Nuclear protein sorting regulates nucleus composition and gene expression, crucial for determining the fate of a eukaryotic cell. Hence, the entry and exit of molecules across the nuclear envelope is a tightly controlled process. Nuclear protein sorting can be inhibited by one of the following ways: 1) masking cargo signal sequences, 2) modifying the nuclear receptor's affinity for cargo, 3) controlling the nuclear pore size, 4) retaining the cargo during its transit to the cytosol or the...
NF-kB-dependent Signaling Pathway02:26

NF-kB-dependent Signaling Pathway

The transcription factor NF-κB was discovered in 1986 in the lab of Nobel laureate Professor David Baltimore, for its interaction with the immunoglobulin light chain enhancer in B-cells. After more than three decades of study, it is now evident that NF-κB regulates the expression of over 100 genes. Most of these genes play an essential role in the innate and adaptive immune responses as well as the inflammatory responses of animals.
NF-κB-dependent Signaling Mechanism
The heterodimer of NF-κB...
Co-activators and Co-repressors02:04

Co-activators and Co-repressors

Gene transcription is regulated by the synergistic action of several proteins that form a complex at a gene regulatory site. This is observed in eukaryotes, where the regulation of gene expression is a complex process. Regulatory proteins in eukaryotes can broadly be classified into two types – regulators that bind directly to specific DNA sequences and co-regulators that associate with regulatory proteins but cannot directly bind to the DNA. These co-regulators are further divided into...
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.
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...