Related Experiment Video

Updated: Mar 7, 2026

Generation of a RIP1 Knockout U937 Cell Line Using the CRISPR-Cas9 System
08:15

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

Published on: April 11, 2025

991

RIP1 autophosphorylation is promoted by mitochondrial ROS and is essential for RIP3 recruitment into necrosome

Yingying Zhang1, Sheng Sean Su1, Shubo Zhao1

  • 1State Key Laboratory of Cellular Stress Biology, Innovation Center for Cell Signaling Network, School of Life Sciences, Xiamen University, Xiamen, Fujian 361005, China.

Nature Communications
|February 9, 2017
PubMed

Insights

Reactive oxygen species (ROS) activate receptor interacting protein 1 (RIP1) kinase activity through cysteine sensing, promoting necroptosis. This ROS-induced RIP1 autophosphorylation at S161 is crucial for necrosome formation and cell death.

Area of Science:

  • Cellular biology
  • Molecular mechanisms of cell death
  • Signal transduction pathways

Background:

  • Necroptosis, a programmed cell death, is implicated in various pathologies.
  • Tumor necrosis factor-alpha (TNF) initiates necroptosis, with mitochondrial reactive oxygen species (ROS) and receptor-interacting protein 1 (RIP1) kinase activity being essential.
  • The precise roles of ROS and RIP1 phosphorylation targets in necroptosis remain unclear.

Purpose of the Study:

  • To elucidate the mechanism by which ROS regulate RIP1 kinase activity in TNF-induced necroptosis.
  • To identify the specific phosphorylation site on RIP1 crucial for necroptosis.
  • To understand the functional consequences of RIP1 phosphorylation in necrosome formation.

Main Methods:

  • Investigated the role of cysteine residues in RIP1 for ROS sensing.
  • Utilized biochemical assays to detect RIP1 autophosphorylation.
  • Assessed the necessity of RIP1 kinase activity and S161 phosphorylation for necrosome formation and necroptosis induction.
  • Examined the interplay between ROS, RIP1, and RIP3 in a feedback loop.

Main Results:

  • Identified three critical cysteine residues in RIP1 essential for sensing ROS.
  • Demonstrated that ROS activate RIP1 autophosphorylation specifically at serine 161 (S161).
  • Confirmed that RIP1 kinase activity's primary role in necroptosis is S161 autophosphorylation, which facilitates RIP1-RIP3 complex (necrosome) formation.
  • Established that ROS function in a positive feedback loop involving RIP3 to ensure robust necroptosis.

Conclusions:

  • RIP1 senses ROS via critical cysteines, leading to S161 autophosphorylation and activation of necroptosis.
  • RIP1 S161 phosphorylation is a key event for necrosome assembly and execution of necroptosis.
  • ROS-mediated positive feedback involving RIP1 and RIP3 amplifies the necroptotic signal, ensuring effective cell death induction.

More Related Videos

Tyramide Signal Amplification for the Immunofluorescent Staining of ZBP1-Dependent Phosphorylation of RIPK3 and MLKL After HSV-1 Infection in Human Cells09:15

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
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...
7.0K
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...
3.1K
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...
4.8K
Translocation of Proteins into the Mitochondria01:19

Translocation of Proteins into the Mitochondria

Mitochondrial precursors are translocated to the internal subcompartments via independent mechanisms involving distinct protein machineries called translocases.
Sorting of outer membrane proteins:
Mitochondrial outer membrane proteins are of two types: the transmembrane, beta-barrel porins, and the membrane-anchored, alpha-helical proteins. Beta-barrel porin precursors are translocated by the TOM complex and inserted into the outer mitochondrial membrane by the SAM complex. In contrast,...
13.6K
PI3K/mTOR/AKT Signaling Pathway01:22

PI3K/mTOR/AKT Signaling Pathway

The mammalian target of rapamycin  (mTOR) is a serine/threonine kinase that regulates growth, proliferation, and cell survival in response to hormones, growth factors, or nutrient availability. This kinase exists in two structurally and functionally distinct forms: mTOR complex 1  (mTORC1) and mTOR complex 2  (mTORC2). The first form (mTORC1) is composed of a rapamycin-sensitive Raptor and proline-rich Akt substrate, PRAS40. In contrast,  mTORC2 consists of a...
6.0K
MAPK Signaling Cascades01:07

MAPK Signaling Cascades

Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...
8.9K