Oxidative stress and mitochondrial dysfunction are centrally involved in the etiology of several diseases and in the normal process of aging. The protein DJ-1 is an oxidative stress response protein whose absence or dysregulation has been implicated in parkinsonism, cancer, and stroke. DJ-1 can robustly protect cells against multiple forms of oxidative stress and thereby confer protection against degeneration that can lead to disease. The molecular mechanism(s) of DJ-1's action, however, remains unclear. DJ-1 contains a functionally essential cysteine residue whose oxidation is hypothesized to regulate its cytoprotective function. We will investigate the mechanism by which DJ-1 senses and responds to oxidative stress by accomplishing three specific aims.
The first aim will investigate the role of DJ-1 cysteine oxidation in the protection against oxidative stress in the Drosophila animal model system. We will combine X-ray crystallography, biochemistry, and Drosophila genetics to establish a powerful animal model for the redox regulation of DJ-1 function.
The second aim will determine the structure-function relationships for an established mRNA binding activity of DJ-1. The results will be used to test the hypothesis that conserved structural features near the oxidized cysteine integrate the RNA binding and redox sensing functions of DJ-1.
The third aim will use a prokaryotic model system to investigate the evolutionarily conserved mechanism of DJ-1 protective function. The results will be used to test existing hypotheses about the conservation of regulatory cysteine oxidation in DJ-1 function as well as discover new functions for DJ-1. In total, the proposed research will provide a comprehensive molecular basis for understanding the oxidative regulation and pathogenic disruption of DJ-1 function. Ultimately, the results of this research will be used to design a new generation of therapeutics that enhance the protective function of DJ-1 in vulnerable cell types.
Oxidative stress and mitochondrial dysfunction are centrally involved in several human diseases. Major recent advances have identified DJ-1 as a protein that confers robust protection against oxidative stress. The precise biochemical function of DJ-1, however, remains uncertain. The long-term goal of this proposal is to determine the biochemical functions of DJ-1 that confer protection against oxidative stress and with the goal of developing therapies that improve the protective function of DJ-1.
|Catazaro, Jonathan; Andrews, Tessa; Milkovic, Nicole M et al. (2017) 15N CEST data and traditional model-free analysis capture fast internal dynamics of DJ-1. Anal Biochem 542:24-28|
|Johnson, William M; Golczak, Marcin; Choe, Kyonghwan et al. (2016) Regulation of DJ-1 by Glutaredoxin 1 in Vivo: Implications for Parkinson's Disease. Biochemistry 55:4519-32|
|Milkovic, Nicole M; Catazaro, Jonathan; Lin, Jiusheng et al. (2015) Transient sampling of aggregation-prone conformations causes pathogenic instability of a parkinsonian mutant of DJ-1 at physiological temperature. Protein Sci 24:1671-85|
|Hasim, Sahar; Hussin, Nur Ahmad; Alomar, Fadhel et al. (2014) A glutathione-independent glyoxalase of the DJ-1 superfamily plays an important role in managing metabolically generated methylglyoxal in Candida albicans. J Biol Chem 289:1662-74|
|Wilson, Mark A (2014) Metabolic role for yeast DJ-1 superfamily proteins. Proc Natl Acad Sci U S A 111:6858-9|
|Wan, Qun; Kovalevsky, Andrey Y; Wilson, Mark A et al. (2014) Preliminary joint X-ray and neutron protein crystallographic studies of ecDHFR complexed with folate and NADP+. Acta Crystallogr F Struct Biol Commun 70:814-8|
|Keedy, Daniel A; van den Bedem, Henry; Sivak, David A et al. (2014) Crystal cryocooling distorts conformational heterogeneity in a model Michaelis complex of DHFR. Structure 22:899-910|
|Wan, Qun; Bennett, Brad C; Wilson, Mark A et al. (2014) Toward resolving the catalytic mechanism of dihydrofolate reductase using neutron and ultrahigh-resolution X-ray crystallography. Proc Natl Acad Sci U S A 111:18225-30|
|Prahlad, Janani; Hauser, David N; Milkovic, Nicole M et al. (2014) Use of cysteine-reactive cross-linkers to probe conformational flexibility of human DJ-1 demonstrates that Glu18 mutations are dimers. J Neurochem 130:839-53|
|Furt, Fabienne; Allen, William J; Widhalm, Joshua R et al. (2013) Functional convergence of structurally distinct thioesterases from cyanobacteria and plants involved in phylloquinone biosynthesis. Acta Crystallogr D Biol Crystallogr 69:1876-88|
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