Nuclear translocation of DJ-1 during oxidative stress-induced neuronal cell death

Free Radic Biol Med. 2012 Aug 15;53(4):936-50. doi: 10.1016/j.freeradbiomed.2012.05.035. Epub 2012 Jun 7.

Abstract

Loss-of-function mutations in the PARK7/DJ-1 gene cause early onset autosomal-recessive Parkinson disease. DJ-1 has been implicated in protection of neurons from oxidative stress and in regulation of transcriptional activity. However, whether there is a relationship between the subcellular localization of DJ-1 and its function remains unknown. Therefore, we examined the subcellular localization of DJ-1 during dopaminergic neurodegeneration induced by various insults. Immunoblotting and immunocytochemistry showed that the nuclear pool of DJ-1 dramatically increased in both MN9D dopaminergic neuronal cells and primary cultures of mesencephalic dopaminergic neurons after 6-hydroxydopamine (6-OHDA) treatment. This was paralleled by a corresponding decrease in its cytosolic level, indicating drug-induced nuclear translocation of DJ-1. The same phenomenon was detected in other cell death paradigms induced by pro-oxidants including hydrogen peroxide and cupric chloride. Consequently, cotreatment with the antioxidant N-acetyl-l-cysteine blocked the translocation of DJ-1 into the nucleus. However, mutation at cysteine 106 had no effect on the translocation of DJ-1 into the nucleus, suggesting that reactive oxygen species-mediated downstream signaling and/or modifications other than oxidative modification are involved in its nuclear translocation. Ectopic expression of nucleus localization signal (NLS)-tagged DJ-1 prevented cell death from 6-OHDA. We investigated whether nuclear DJ-1 was involved in transcriptional regulation and found that DJ-1 was localized in promyelocytic leukemia bodies, and this localization increased upon 6-OHDA treatment. We also confirmed that binding of DJ-1 and promyelocytic leukemia bodies indeed increased after 6-OHDA treatment. Consequently, expression levels of acetylated p53 and PUMA were downregulated in cells overexpressing DJ-1 or NLS-tagged DJ-1. Taken together, our data suggest that nuclear translocation of DJ-1 may protect neurons from cell death after oxidative stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcysteine / pharmacology
  • Active Transport, Cell Nucleus
  • Animals
  • Antioxidants / pharmacology
  • Apoptosis Regulatory Proteins / metabolism
  • Apoptosis*
  • Caspase 3 / metabolism
  • Cells, Cultured
  • Copper / pharmacology
  • Dopaminergic Neurons / drug effects
  • Dopaminergic Neurons / metabolism
  • Dopaminergic Neurons / physiology*
  • Humans
  • Intracellular Signaling Peptides and Proteins / metabolism*
  • Mice
  • Oncogene Proteins / metabolism*
  • Oxidants / pharmacology
  • Oxidative Stress*
  • Oxidopamine / pharmacology
  • Primary Cell Culture
  • Protein Deglycase DJ-1
  • Proto-Oncogene Proteins / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Reactive Oxygen Species / metabolism

Substances

  • Antioxidants
  • Apoptosis Regulatory Proteins
  • BBC3 protein, human
  • Intracellular Signaling Peptides and Proteins
  • Oncogene Proteins
  • Oxidants
  • Proto-Oncogene Proteins
  • Reactive Oxygen Species
  • Copper
  • Oxidopamine
  • PARK7 protein, human
  • Protein Deglycase DJ-1
  • CASP3 protein, human
  • Caspase 3
  • cupric chloride
  • Acetylcysteine