Related Experiment Video

Updated: Mar 16, 2026

Tyramide Signal Amplification for the Immunofluorescent Staining of ZBP1-Dependent Phosphorylation of RIPK3 and MLKL After HSV-1 Infection in Human Cells
09:15

Tyramide Signal Amplification for the Immunofluorescent Staining of ZBP1-Dependent Phosphorylation of RIPK3 and MLKL After HSV-1 Infection in Human Cells

Published on: October 20, 2022

2.9K

Coordinated ubiquitination and phosphorylation of RIP1 regulates necroptotic cell death

M Cristina de Almagro1, Tatiana Goncharov1, Anita Izrael-Tomasevic2

  • 1Department of Early Discovery Biochemistry, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.

Cell Death and Differentiation
|August 13, 2016
PubMed

Insights

Necroptosis, a regulated cell death pathway, is controlled by RIP1 ubiquitination and phosphorylation within the necrosome. This dual regulation is vital for necroptotic signaling and preventing diseases like inflammatory bowel disease.

Area of Science:

  • Cellular biology
  • Molecular mechanisms of cell death
  • Biochemistry

Background:

  • Regulated cell death is critical for tissue homeostasis and disease prevention.
  • Necroptosis, a caspase-independent cell death, is implicated in inflammatory bowel disease and ischemia-reperfusion injury.
  • Activation of necroptosis involves RIP1 and RIP3 kinases, leading to inflammatory cell death.

Purpose of the Study:

  • To investigate the role of RIP1 ubiquitination in necroptotic signaling.
  • To identify ubiquitination sites on RIP1 during necroptosis.
  • To understand the interplay between RIP1 phosphorylation and ubiquitination in regulating necroptosis.

Main Methods:

  • Immunoaffinity enrichment coupled with mass spectrometry to profile RIP1 ubiquitination.
  • Site-directed mutagenesis of RIP1 to assess the impact on necroptosis.
  • Analysis of RIP1 and RIP3 complex assembly in the necrosome.

Main Results:

  • Necroptotic cell death is regulated by RIP1 ubiquitination in the necrosome, in addition to phosphorylation.
  • RIP1 ubiquitination requires RIP1 kinase activity but not RIP3 or MLKL.
  • Mutation of a key RIP1 ubiquitination site impaired necroptosis, RIP1 ubiquitination/phosphorylation, and necrosome assembly.
  • RIP1 ubiquitination was observed in injured kidneys, suggesting a physiological role in ischemia-reperfusion injury.

Conclusions:

  • RIP1 ubiquitination is a crucial regulator of necroptotic signaling and cell death.
  • Coordinated RIP1 phosphorylation and ubiquitination within the necrosome are essential for maintaining RIP1 kinase activity.
  • These findings highlight RIP1 ubiquitination as a potential therapeutic target for necroptosis-related diseases.

More Related Videos

Generation of a RIP1 Knockout U937 Cell Line Using the CRISPR-Cas9 System08:15

Generation of a RIP1 Knockout U937 Cell Line Using the CRISPR-Cas9 System

Published on: April 11, 2025

1.0K
Characterization of MLKL-mediated Plasma Membrane Rupture in Necroptosis08:55

Characterization of MLKL-mediated Plasma Membrane Rupture in Necroptosis

Published on: August 7, 2018

11.4K

Related Concept Videos

Necrosis01:16

Necrosis

Necrosis is considered as an “accidental” or unexpected form of cell death that ends in cell lysis. The first noticeable mention of “necrosis” was in 1859 when Rudolf Virchow used this term to describe advanced tissue breakdown in his compilation titled “Cell Pathology”.
Morphological Manifestations of Necrosis
Necrotic cells show different types of morphological appearance depending on the type of tissue and infection. In coagulative necrosis, cells become...
7.1K
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...
3.2K
Autophagic Cell Death01:18

Autophagic Cell Death

Christian de Duve discovered “autophagy,” a process in which cellular components are engulfed by membrane-bound organelles called autophagosomes. The autophagosomes then fuse with lysosomes to digest the enclosed contents. Autophagy is generally activated in cells to prevent cell death. However, cell death is triggered when the damage is beyond repair.
Autophagy and Apoptosis
Autophagy can activate apoptosis. In normal conditions, the autophagy activating protein Beclin-1 and...
4.9K
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...
9.1K
Regulated Protein Degradation02:58

Regulated Protein Degradation

3.3K
The Extrinsic Apoptotic Pathway01:17

The Extrinsic Apoptotic Pathway

The extrinsic apoptotic pathway is initiated when extracellular death-inducing signals, such as specific cytokines, activate the death receptors expressed on the cell surface. The immune cells involved in this pathway are natural killer cells (NK cells) and cytotoxic T-lymphocytes. NK cells are critical in innate immune response, while cytotoxic T-lymphocytes are associated with adaptive immune response. These cells recognize specific receptors expressed on the altered cells and activate...
9.2K