Effect of RNF113A deficiency on oxidative stress-induced NRF2 pathway

Abstract

The ring finger protein 113A (RNF113A) serves as an E3 ubiquitin ligase and a subunit of the spliceosome. Mutations in the RNF113A gene are associated with X-linked trichothiodystrophy (TTD). However, the cellular roles of RNF113A remain largely unknown. In this study, we performed transcriptome profiling of RNF113A knockout (KO) HeLa cells using RNA sequencing and revealed the upregulation of NRF2 pathway-associated genes. Further analysis confirmed that the KO of RNF113A promotes nuclear localization of the NRF2 protein and elevates the mRNA levels of NRF2 target genes. RNF113A KO cells showed high levels of intracellular reactive oxygen species (ROS) and decreased resistance to cell death following H 2 O 2 treatment. Additionally, RNF113A KO cells more sensitively formed stress granules (SGs) under arsenite-induced oxidative stress. Moreover, RNF113A KO cells exhibited a decrease in glutathione levels, which could be attributed to a reduction in GLUT1 expression levels, leading to decreased glucose uptake reactions and lower intracellular glucose levels. These alterations potentially caused a reduction in ROS scavenging activity. Taken together, our findings suggest that the loss of RNF113A promotes oxidative stress-mediated activation of the NRF2 pathway, providing novel insights into RNF113A-associated human diseases.

Establishment of a stress granule reporter system for evaluating in vitro colon toxicity

“…We also confirmed that these cells could potentially be used as an in vitro colon toxicity monitoring system based on real-time reporting of stress granule formation. Stress granule formation is a highly sensitive and acute defense mechanism that responds to a wide range of cellular stress stimuli, including oxidative stress, ER stress, hypoxia, osmotic stress, unfolded protein responses, and viral infections (Cho et al 2024 ; Jung et al 2023 ; Kang et al 2018 ; Wang et al 2020 ; Wang et al 2021 ; Zeng et al 2020 ). These features may be a powerful advantage in our established cell lines and could contribute to the sensitive reporting of cellular toxicity from a broad range of cellular stresses.…”