The effects of DJ-1 and A20 family members on inflammation and oxidative response Public Deposited

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Last Modified
  • March 21, 2019
Creator
  • McNally, Richard Sean
    • Affiliation: School of Medicine, Department of Microbiology and Immunology
Abstract
  • Dysregulation of the balance between cell survival and cell death is seen in the loss of dopaminergic neurons in Parkinson's disease and the inappropriate cell survival in cancer. Changes in DJ-1 expression are associated with both Parkinson's disease and cancer suggesting an important role of DJ-1 in this balance. Our lab has reported that DJ-1 positively regulates the antioxidant response transcription factor Nrf2 protecting cells from ROS and toxic insults. Although this work revealed a mechanism for DJ-1's effect on cell survival, we could not find a direct role of DJ-1 as DJ-1 did not bind any proteins in the Nrf2 pathway. Using Q-TOF mass spectrometry, we found that BBS1, CLCF1, MTREF, and Cezanne interact with DJ-1. We focused our attention on the de-ubiquitinating enzyme Cezanne due to its known inhibition of the cancer-associated transcription factor NF-kB. We confirmed the interaction between DJ-1 and Cezanne and found that it inhibits Cezanne's de-ubiquitinating activity. Using shRNA against DJ-1 and Cezanne, we found that DJ-1 positively regulates NF-kB nuclear localization and promotes cell survival while Cezanne has reciprocal effects. Using microarray, we identified IL-8 and ICAM-1 as downstream targets of DJ-1 and Cezanne's effects on NF-kB. This finding was confirmed at the RNA and protein levels using real-time PCR, ELISA, and immunoblot. Finally, we identified the presence of this pathway in a primary cell type. This work is the first to identify a link between DJ-1 and NF-kB representing a novel mechanism by which DJ-1 can control cell survival. During this research, we also identified a putative mechanism of crosstalk between the Nrf2 and NF-kB pathways as two well known inhibitors of these pathways, Keap1 and A20, were found to interact. Unfortunately, the consequences of the Keap1/A20 interaction have not been elucidated due to the complex interplay between the Nrf2 and NF-kB pathways. Taken as a whole, this work reports that DJ-1 is one of the few proteins or stimuli known to enhance cell survival through the activation of both Nrf2 and NF-kB while also identifying a novel mechanism of crosstalk between these pathways with implications in both Parkinson's disease and cancer.
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  • In Copyright
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  • "... in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Department of Microbiology and Immunology."
Advisor
  • Ting, Jenny P.-Y.
Degree granting institution
  • University of North Carolina at Chapel Hill
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Place of publication
  • Chapel Hill, NC
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  • Open access
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