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Expression profiles of genes in DJ-1-knockdown and L 166 P DJ-1 mutant cells
Hiromi Nishinaga1, Kazuko Takahashi-Niki, Takahiro Taira
1Graduate School of Pharmaceutical Sciences, Hokkaido University, Sapporo 060-0812, Japan.
Abstract:
DJ-1 is a novel oncogene and a causative gene for the familial form of Parkinson's disease (PD). DJ-1 has been shown to play roles in anti-oxidative stress by eliminating reactive oxygen species and in transcriptional regulation of genes. Loss of these functions of DJ-1 is thought to trigger the onset of PD. In this study, to identify genes for which expressions are regulated by DJ-1, DNA microarray analyses were carried out using two mouse NIH3T3 cell lines, DJ-1-knockdown cells and cells harboring an exogenously added L 166 P DJ-1 mutant found in PD patients. In both cell lines, drastic changes in expressions of genes, including genes related to stress, apoptosis, oxidative stress and neurotoxicity, were observed and changes in expressions were confirmed by RT-PCR. Of the genes identified, expression level of the extracellular superoxide dismutase (SOD 3) gene was found to decrease in DJ-1-knockdown cells, while expressions of SOD 1 and SOD 2 genes did not change. Furthermore, expression of the tau gene, a gene whose product gives cells neurotoxicity by aggregation, was found to increase at its promoter level in L 166 P DJ-1 cells. These findings suggest that DJ-1 regulates expressions of genes for which functions are thought to be related to cell death or neurodegeneration.
Insights
DJ-1, a gene linked to Parkinson's disease (PD), regulates genes involved in cell death. This study identified specific gene expression changes in DJ-1-deficient cells, offering insights into PD pathogenesis.
Area of Science:
- Molecular Biology
- Neuroscience
- Genetics
Background:
- DJ-1 is implicated as an oncogene and a causative gene in familial Parkinson's disease (PD).
- DJ-1's known functions include anti-oxidative stress and transcriptional regulation, with loss of these functions potentially triggering PD.
- Understanding DJ-1's regulatory targets is crucial for elucidating PD mechanisms.
Purpose of the Study:
- To identify genes whose expression is regulated by DJ-1.
- To investigate the impact of DJ-1 deficiency and a specific PD-associated mutant (L166P) on gene expression profiles.
Main Methods:
- DNA microarray analysis was performed on DJ-1-knockdown mouse NIH3T3 cells and cells expressing the L166P DJ-1 mutant.
- Gene expression changes were confirmed using Reverse Transcription Polymerase Chain Reaction (RT-PCR).
Main Results:
- Significant alterations in the expression of genes related to stress, apoptosis, oxidative stress, and neurotoxicity were observed.
- Extracellular superoxide dismutase (SOD3) gene expression decreased in DJ-1-knockdown cells.
- Tau gene expression increased at the promoter level in L166P DJ-1 mutant cells, suggesting a role in neurotoxicity.
Conclusions:
- DJ-1 plays a regulatory role in genes associated with cell death and neurodegeneration.
- The findings provide molecular insights into how DJ-1 dysfunction contributes to Parkinson's disease.
- Specific gene targets like SOD3 and tau are identified as being modulated by DJ-1.
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