Related Experiment Video
Updated: Jan 4, 2026
Monitoring Hippo Signaling Pathway Activity Using a Luciferase-based Large Tumor Suppressor LATS Biosensor
Published on: September 13, 2018
Neural precursor cell-expressed, developmentally down-regulated 4 (NEDD4) regulates hydrogen peroxide-induced cell
Seon-Ae Jeon1, Dong Wook Kim1, Je-Yoel Cho1
1Department of Veterinary Biochemistry, BK21 Plus and Research Institute for Veterinary Science, School of Veterinary Medicine, Seoul National University, Seoul, South Korea.
Abstract:
E3 ubiquitin ligases are involved in the regulation of oxidative stress-induced cell death. In this study, we investigated the role of neural precursor cell-expressed, developmentally down-regulated protein 4 (NEDD4) in regulation of hydrogen peroxide (H2O2)-induced cell proliferation and apoptosis in human bone marrow-derived stem cells (hBMSCs). Cell proliferation was increased in low doses of H2O2 (10-4 to 10-2 μM), whereas sublethal concentrations of H2O2 (>200 μM) induced apoptosis. A chromatin immunoprecipitation assay identified that recruitment of NF-κB onto the promoter region of NEDD4 mediated H2O2-induced NEDD4 expression. The increase of NEDD4 expression by H2O2 induced translocation of yes-associated protein (YAP) into the nucleus by decreasing the stability of large tumor suppressor kinase (LATS). Thus, the phosphorylation of serine 127 residue of YAP by LATS upstream kinase is decreased and thereby increased the transcriptional activity of YAP. The mRNA expression levels of catalase and manganese superoxide dismutase, which are well-known targets of YAP, were increased by H2O2 treatment but down-regulated by NEDD4 silencing using a specific small interfering RNA targeting NEDD4 (siNEDD4). H2O2-induced scavenging capacity of reactive oxygen species was also decreased by siNEDD4 in hBMSCs. Finally, hBMSC differentiation into osteoblast was decreased by siNEDD4 but reverted by reintroduction of the S127A mutant construction of YAP. Taken together, these results indicate that NEDD4 regulates H2O2-induced alteration of cell status through regulation of the Hippo signaling pathway.-Jeon, S.-A., Kim, D. W., Cho, J.-Y. Neural precursor cell-expressed, developmentally down-regulated 4 (NEDD4) regulates hydrogen peroxide-induced cell proliferation and death through inhibition of Hippo signaling.
Insights
Neural precursor cell-expressed, developmentally down-regulated protein 4 (NEDD4) regulates hydrogen peroxide (H2O2)-induced cell proliferation and apoptosis in stem cells. NEDD4 impacts the Hippo signaling pathway, affecting cell fate and antioxidant capacity.
Area of Science:
- Cell Biology
- Stem Cell Research
- Molecular Biology
Background:
- E3 ubiquitin ligases play a crucial role in regulating oxidative stress-induced cell death.
- Hydrogen peroxide (H2O2) at varying concentrations can induce either cell proliferation or apoptosis in human bone marrow-derived stem cells (hBMSCs).
- The specific role of neural precursor cell-expressed, developmentally down-regulated protein 4 (NEDD4) in H2O2-induced cellular responses requires elucidation.
Purpose of the Study:
- To investigate the function of NEDD4 in modulating H2O2-induced cell proliferation and apoptosis in hBMSCs.
- To elucidate the molecular mechanisms by which NEDD4 influences cellular responses to oxidative stress, particularly its interaction with the Hippo signaling pathway.
Main Methods:
- Chromatin immunoprecipitation (ChIP) assay to identify transcription factor binding to the NEDD4 promoter.
- Small interfering RNA (siRNA) for NEDD4 silencing (siNEDD4) to assess its functional impact.
- Analysis of yes-associated protein (YAP) nuclear translocation, phosphorylation, and transcriptional activity.
- Measurement of mRNA expression levels of antioxidant enzymes (catalase, manganese superoxide dismutase) and reactive oxygen species (ROS) scavenging capacity.
- Assessment of hBMSC differentiation into osteoblasts.
Main Results:
- H2O2 treatment increased NEDD4 expression, mediated by NF-κB recruitment to the NEDD4 promoter.
- Increased NEDD4 expression promoted YAP nuclear translocation by destabilizing LATS, leading to decreased YAP phosphorylation and enhanced transcriptional activity.
- NEDD4 silencing reduced H2O2-induced expression of YAP target genes (catalase, manganese superoxide dismutase) and impaired ROS scavenging capacity.
- NEDD4 silencing inhibited osteogenic differentiation of hBMSCs, an effect reversible by introducing a YAP mutant.
Conclusions:
- NEDD4 is a key regulator of H2O2-induced cellular responses in hBMSCs, influencing proliferation, apoptosis, and differentiation.
- NEDD4 exerts its regulatory function through the inhibition of the Hippo signaling pathway, impacting YAP activity.
- These findings highlight NEDD4 as a potential therapeutic target for conditions involving oxidative stress and stem cell dysfunction.
Related Concept Videos
02:33Hedgehog Signaling Pathway
01:24Peroxisomes
01:24Regulation of Angiogenesis and Blood Supply
01:12Role Of Notch Signalling In Intestinal Stem Cell Renewal
Direct cell-to-cell contact is needed for the activation of Notch signaling. The signal is initiated when a notch ligand binds to a receptor on an adjacent cell, also...
01:19Interactions Between Signaling Pathways
Convergence and divergence, and cross-talk between signaling pathways
Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...
03:14Notch Signaling Pathway
The Notch gene came into the limelight in 1914 after the discovery that its mutation in Drosophila melanogaster leads to a serrated (or "notched") wing margin phenotype. It was not...