Oxidative Stress Induces Neuronal Apoptosis Through Suppressing Transcription Factor EB Phosphorylation at Ser467

Abstract

Background/Aims: This study determined the role and mechanism of action of transcription factor EB (TFEB) in H₂O₂-induced neuronal apoptosis. Methods: SH-SY5Y cells were treated with Akt inhibitor/activator and different concentrations of H₂O₂. Cell apoptosis was detected by flow cytometric analysis. Akt and TFEB phosphorylation and PARP cleavage were determined by Western blotting. HEK293T cells were transfected with different truncated TFEB mutants and HA-Akt-WT; SH-SY5Y cells were transfected with Flag-vector, Flag-TFEB, Flag-TFEB-S467A or Flag-TFEB-S467D; and TFEB interaction with Akt was determined by co-immunoprecipitation and GST pull-down assays. Results: A low concentration of H₂O₂ induces TFEB phosphorylation at Ser467 and nuclear translocation, facilitating neuronal survival, whereas a high concentration of H₂O₂ promotes SH-SY5Y cell apoptosis via suppressing TFEB Ser467 phosphorylation and nuclear translocation. The TFEB-S467D mutant is more easily translocated into the nucleus than the non-phosphorylated TFEB-S467A mutant. Further, Akt physically binds to TFEB via its C-terminal tail interaction with the HLH domain of TFEB and phosphorylates TFEB at Ser467. Mutation of TFEB-Ser467 can prevent the phosphorylation of TFEB by Akt, preventing inhibition of oxidative stress-induced apoptosis. Conclusions: Oxidative stress induces neuronal apoptosis through suppressing TFEB phosphorylation at Ser467 by Akt, providing a novel therapeutic strategy for neurodegenerative diseases.

Hydrogen Inhalation Attenuates Oxidative Stress Related Endothelial Cells Injury After Subarachnoid Hemorrhage in Rats

“…Reactive oxygen species generated by mitochondrial dysfunction play a significant role in oxidative stress to various types of cells in the brain including microglia, astrocytes, neurons, and endothelial cells, which has been related to inflammation, apoptosis and neurological deficits after SAH (Ayer and Zhang, 2008;Sorce and Krause, 2009;Skowronska and Albrecht, 2013;Sahebkar et al, 2018;Su et al, 2018). The NLRP3 inflammasome composed of NLRP3, ASC and caspase-1, is responsible for the maturation and secretion of proinflammatory cytokines IL-1β and IL-18.…”

Rh-CSF1 Attenuates Oxidative Stress and Neuronal Apoptosis via the CSF1R/PLCG2/PKA/UCP2 Signaling Pathway in a Rat Model of Neonatal HIE

“…Neurons were more vulnerable to ischemia and chemical damage in CSF1-deficient mice [ 58 ]. Following HI, OS was significantly increased which are associated with neuroinflammation, mitochondrial dysfunction, glial/axonal mutagenesis, and synaptic transmission disorders, all contributing to progressive loss of neurons including neuronal apoptosis [ 13 , 59 ]. Our study confirmed that there are increased cellular OS markers MitoSOX and 8-OHdG associated with more FJC-/TUNEL-positive degenerated and apoptotic neurons within the perilesion brain regions of HI rat pups.…”

Evaluating and modulating TFEB in the control of autophagy: toward new treatments in CNS disorders

“…These ROS irreversibly oxidize DNA and cellular biomolecules, which creates an additional source of damage in biological systems [37]. Oxidative stress is also more pronounced in neurodegenerative disorders such as Parkinson's disease (PD), where modulation of TFEB has been proposed to reduce neuronal cell death [38,39]. Low concentrations of H 2 O 2 result in the phosphorylation of TFEB by Akt on Ser467, which promotes its nuclear translocation, thus facilitating neuronal survival.…”

“…Low concentrations of H 2 O 2 result in the phosphorylation of TFEB by Akt on Ser467, which promotes its nuclear translocation, thus facilitating neuronal survival. On the other hand, higher concentrations of H 2 O 2 promote apoptosis in SH-SY5Y TFEB, a new target in CNS disorder cells by suppressing TFEB phosphorylation and nuclear translocation [39].…”