SMARCB1 Promotes Ubiquitination and Degradation of NR4A3 via Direct Interaction Driven by ROS in Vascular Endothelial Cell Injury

Oxid Med Cell Longev. 2020 Oct 23;2020:2048210. doi: 10.1155/2020/2048210. eCollection 2020.

Abstract

Nuclear receptor subfamily 4 group A member 3 (NR4A3) protects the vascular endothelial cell (VEC) against hypoxia stress, whose expression is primarily reported to be governed at a transcriptional level. However, the regulation of NR4A3 in the protein level is largely unknown. Here, we report that NR4A3 protein abundance is decreased immensely in VEC injury induced by reoxygenation after oxygen-glucose deprivation (OGD-R), which is significantly blocked by the administration of the antioxidative steroid TRIOL. Moreover, the notable improvement of NR4A3 and the alleviation of pulmonary endothelial barrier hyperpermeability induced by acute hypobaric hypoxia in cynomolgus monkeys are also observed after TRIOL administration. The overproduction of reactive oxygen species (ROS) decreases NR4A3 protein abundance in VEC under OGD-R condition, which is reversed by TRIOL and N-acetylcysteine (NAC). TRIOL dose-dependently increases the NR4A3 protein level by inhibiting ubiquitination and ubiquitin proteasome system- (UPS-) mediated degradation rather than promoting its transcription. Using yeast two-hybrid screening, we further identify the interaction between NR4A3 and SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily B member 1 (SMARCB1), and the DNA-binding domain of NR4A3 is required for this interaction. Knockdown of SMARCB1 reduces ubiquitination and degradation of NR4A3, suggesting the proubiquitylation effect of this interaction which is enhanced by ROS in VEC injury induced by OGD-R. In summary, our study here for the first time reveals a posttranslational regulation in SMARCB1-mediated NR4A3 protein degradation which is driven by ROS, providing further understanding of the impaired regulation of NR4A3-mediated prosurvival pathways under pathological condition in VEC.

MeSH terms

  • Animals
  • DNA-Binding Proteins / metabolism*
  • Endothelium, Vascular / injuries*
  • Endothelium, Vascular / metabolism
  • Endothelium, Vascular / pathology
  • Human Umbilical Vein Endothelial Cells / metabolism*
  • Human Umbilical Vein Endothelial Cells / pathology
  • Humans
  • Hypoxia / metabolism*
  • Lung / metabolism*
  • Lung / pathology
  • Macaca fascicularis
  • Proteolysis*
  • Reactive Oxygen Species*
  • Receptors, Steroid / metabolism*
  • Receptors, Thyroid Hormone / metabolism*
  • SMARCB1 Protein / metabolism*
  • Ubiquitination*

Substances

  • DNA-Binding Proteins
  • NR4A3 protein, human
  • Reactive Oxygen Species
  • Receptors, Steroid
  • Receptors, Thyroid Hormone
  • SMARCB1 Protein
  • SMARCB1 protein, human