Redox Regulation of DJ-1 Function

描述（由申请人提供）：氧化应激和线粒体功能障碍在几种疾病的病因学和正常衰老过程中起核心作用。蛋白DJ-1是一种氧化应激反应蛋白，其缺失或失调与帕金森病、癌症和中风有关。DJ-1可以强有力地保护细胞免受多种形式的氧化应激，从而保护细胞免受可能导致疾病的退化。然而，DJ-1作用的分子机制尚不清楚。DJ-1含有一种功能性必需的半胱氨酸残基，其氧化被认为可以调节其细胞保护功能。我们将通过完成三个特定目标来研究DJ-1感知和响应氧化应激的机制。第一个目的是研究DJ-1半胱氨酸氧化在果蝇动物模型系统中抗氧化应激的作用。我们将结合x射线晶体学、生物化学和果蝇遗传学，建立一个强大的动物模型来研究DJ-1功能的氧化还原调控。第二个目标是确定DJ-1 mRNA结合活性的结构-功能关系。该结果将用于验证氧化半胱氨酸附近的保守结构特征整合了DJ-1的RNA结合和氧化还原传感功能的假设。第三个目标是利用原核模型系统研究DJ-1保护功能的进化保守机制。这些结果将用于验证关于DJ-1功能中调节半胱氨酸氧化守恒的现有假设，并发现DJ-1的新功能。总而言之，本研究将为了解DJ-1功能的氧化调控和致病性破坏提供全面的分子基础。最终，这项研究的结果将用于设计新一代的治疗方法，以增强DJ-1对易感细胞类型的保护功能。

DESCRIPTION (provided by applicant): 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.