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.

Public Health Relevance

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.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM092999-03
Application #
8302424
Study Section
Neural Oxidative Metabolism and Death Study Section (NOMD)
Program Officer
Anderson, Vernon
Project Start
2010-08-01
Project End
2015-07-31
Budget Start
2012-08-01
Budget End
2013-07-31
Support Year
3
Fiscal Year
2012
Total Cost
$267,410
Indirect Cost
$79,310
Name
University of Nebraska Lincoln
Department
Biochemistry
Type
Schools of Earth Sciences/Natur
DUNS #
555456995
City
Lincoln
State
NE
Country
United States
Zip Code
68588
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
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
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
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
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
Lin, Jiusheng; Prahlad, Janani; Wilson, Mark A (2012) Conservation of oxidative protein stabilization in an insect homologue of parkinsonism-associated protein DJ-1. Biochemistry 51:3799-807
Lin, Jiusheng; Nazarenus, Tara J; Frey, Jeanine L et al. (2011) A plant DJ-1 homolog is essential for Arabidopsis thaliana chloroplast development. PLoS One 6:e23731
Wilson, Mark A (2011) The role of cysteine oxidation in DJ-1 function and dysfunction. Antioxid Redox Signal 15:111-22
Lin, Jiusheng; Wilson, Mark A (2011) Escherichia coli thioredoxin-like protein YbbN contains an atypical tetratricopeptide repeat motif and is a negative regulator of GroEL. J Biol Chem 286:19459-69

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