Related Experiment Video

Updated: Jun 15, 2025

Deciphering the Structural Effects of Activating EGFR Somatic Mutations with Molecular Dynamics Simulation

Deciphering the Structural Effects of Activating EGFR Somatic Mutations with Molecular Dynamics Simulation

Published on: May 20, 2020

RNF213-Dependent EGFR and HER2 Activation Regulates Specific Downstream Signaling Pathways in Human Cancer Cells

Intisar M Fouad^1,2, Jungmi Choi¹, Qianying Huang¹

¹Laboratory of Molecular Biosciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan.

Genes to Cells : Devoted to Molecular & Cellular Mechanisms

|June 12, 2025

Abstract:

In this study, we reveal a novel relationship between RNF213, an E3 ubiquitin ligase associated with Moyamoya disease (MMD) and the ubiquitination of both endogenous and pathogenic substrates, and EGFR, the epithelial growth factor receptor involved in cell growth, angiogenesis, and cancer. RNF213 knockdown or knockout in HeLa and A549 cells markedly reduces EGFR phosphorylation at key tyrosine sites following EGF and TGFα stimulation. In RNF213 knockout cells, HER2 phosphorylation, typically activated through heterodimerization with EGFR, and Src recruitment and/or phosphorylation are also diminished. Mutations in the RNF213 RING, RZ finger, or AAA+ domains, including the prevalent R4810K mutation in MMD, consistently reduce EGFR phosphorylation. In vivo, EGF injections increase EGFR and HER2 phosphorylation in WT but not in RNF213 knockout mice. Despite the reduced phosphorylation levels of these tyrosine kinases in knockout cells, the activation of downstream signals such as AKT, ERK1/2, and STAT3 remains unaffected, although phosphorylation of PLCγ, a key mediator of Ca²⁺ release, is selectively reduced by RNF213 knockout. These findings demonstrate that RNF213 modulates EGFR-related pathways and specific downstream signal pathways, possibly affecting physiologic and pathogenic angiogenesis, and may have implications for unraveling the etiology of MMD and for developing cancer therapies that target RNF213.

Insights

RNF213, an E3 ubiquitin ligase linked to Moyamoya disease, modulates epithelial growth factor receptor (EGFR) phosphorylation. This discovery impacts understanding of angiogenesis and cancer, potentially informing new therapies.

Area of Science:

Molecular Biology
Cell Signaling
Biochemistry

Background:

RNF213 is an E3 ubiquitin ligase associated with Moyamoya disease (MMD).
Epidermal Growth Factor Receptor (EGFR) signaling is crucial for cell growth, angiogenesis, and cancer.
The precise role of RNF213 in EGFR signaling pathways remains largely unexplored.

Purpose of the Study:

To investigate the novel relationship between RNF213 and EGFR signaling.
To determine the impact of RNF213 on EGFR phosphorylation and downstream signaling.
To explore the implications for Moyamoya disease and cancer therapeutics.

Main Methods:

RNF213 knockdown and knockout in HeLa and A549 cell lines.
Stimulation with EGF and TGFα to assess EGFR phosphorylation.
Analysis of HER2, Src, AKT, ERK1/2, STAT3, and PLCγ phosphorylation.
In vivo studies using RNF213 knockout mice and EGF injections.

Main Results:

RNF213 deficiency significantly reduces EGFR phosphorylation at key tyrosine sites.
HER2 phosphorylation and Src recruitment are diminished in RNF213 knockout cells.
Specific RNF213 mutations, including R4810K, impair EGFR phosphorylation.
Downstream signaling via AKT, ERK1/2, and STAT3 is unaffected, but PLCγ phosphorylation is reduced.

Conclusions:

RNF213 plays a critical role in modulating EGFR-related signaling pathways.
The findings suggest RNF213 influences angiogenesis and may be implicated in MMD pathogenesis.
Targeting RNF213 could offer a novel therapeutic strategy for MMD and certain cancers.

Read full analysis

Keywords:

EGFR HER2 Moyamoya disease PLCγ RNF213 Src angiogenesis cancer phosphorylation vasculopathy

More Related Videos

Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure

Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure

Published on: August 11, 2017

Identification of EGFR and RAS Inhibitors using Caenorhabditis elegans

Identification of EGFR and RAS Inhibitors using Caenorhabditis elegans

Published on: October 5, 2020

Related Concept Videos

Mitogens and the Cell Cycle

Mitogens and the Cell Cycle

Mitogens and their receptors play a crucial role in controlling the progression of the cell cycle. However, the loss of mitogenic control over cell division leads to tumor formation. Therefore, mitogens and mitogen receptors play an important role in cancer research. For instance, the epidermal growth factor (EGF) - a type of mitogen and its transmembrane receptor (EGFR), decides the fate of the cell's proliferation. When EGF binds to EGFR, a member of the ErbB family of tyrosine kinase...

The Ras Gene

The Ras Gene

The Ras-gene-encoded proteins are regulators of signaling pathways controlling cell proliferation, differentiation, or cell survival. The Ras-gene family in humans constitutes three primary members—the HRas, NRas, and KRas. These genes code for four functionally distinct yet closely related proteins—the HRas, NRas, KRas4A, and KRas4B. The involvement of mutant Ras genes in human cancer was first discovered in 1982 and is among the most common causes of human tumorigenesis.
Ras is a...

mTOR Signaling and Cancer Progression

mTOR Signaling and Cancer Progression

The mammalian target of rapamycin or mTOR protein was discovered in 1994 due to its direct interaction with rapamycin. The protein gets its name from a yeast homolog called TOR. The mTOR protein complex in mammalian cells plays a major role in balancing anabolic processes such as the synthesis of proteins, lipids, and nucleotides and catabolic processes, such as autophagy in response to environmental cues, such as availability of nutrients and growth factors.
The mTOR pathway or the...

NF-κB-dependent Signaling Pathway

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...

MAPK Signaling Cascades

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...

Abnormal Proliferation

Abnormal Proliferation

Under normal conditions, most adult cells remain in a non-proliferative state unless stimulated by internal or external factors to replace lost cells. Abnormal cell proliferation is a condition in which the cell's growth exceeds and is uncoordinated with normal cells. In such situations, cell division persists in the same excessive manner even after cessation of the stimuli, leading to persistent tumors. The tumor arises from the damaged cells that replicate to pass the damage to the...

Articles linked to this work by shared authors, journal, and citation graph.

RNF213 regulates palmitate signaling through AAA+ and RZ-finger domains, a process enhanced by gain-of-function moyamoya disease-associated mutations.

The FEBS journal·2026

Low-dose docetaxel reprograms CAR T cells for enhanced solid tumor immunity.

Journal of experimental & clinical cancer research : CR·2026

Molecular Functions of Ubiquitin-like Modifiers in Bacterial Infection.

Cells·2026

Food-Derived Elastin Peptides Improve Glucose Metabolism and Protect Renal Vasculature in Stroke-Prone Spontaneously Hypertensive Rats Despite Modest Dipeptidyl Peptidase 4 Inhibition.

Nutrients·2026

Delivering living medicines with polysaccharide derivatives for disease treatment.

Journal of controlled release : official journal of the Controlled Release Society·2026

Advanced strategies to enhance the safety, persistence, and efficacy of CAR-T cells in solid tumors.

Frontiers in immunology·2026

See all related articles