Genetic Determinants of Oxygen Toxicity

氧中毒的遗传决定因素

• 批准号: 7471381
• 负责人: Valeria C Culotta
• 金额: $ 32.66万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7471381
• 关键词: Address; Age Factors; Aging; Alleles; Amyotrophic Lateral Sclerosis; Antioxidants; Biochemical; Biochemical Genetics; Biochemistry; Biology; Chromatin; Chromatin Remodeling Factor; Condition; Copper; Cysteine; Cytosol; Disease; Disulfides; Electrons; Enzyme Stability; Enzymes; Family; Fostering; Funding; Genetic; Genetic Determinism; Glutathione; Goals; Health; Human; Investigation; Lead; Light; Mammalian Cell; Metals; Molecular Chaperones; Molecular Genetics; Monitor; Mutagenesis; Mutation; Numbers; Organism; Oxidation-Reduction; Pathway interactions; Play; Positioning Attribute; Post-Translational Protein Processing; Process; Proline; Proteins; RNA Interference; Range; Repression; Research; Research Personnel; Research Proposals; Role; Series; Sirtuins; Structural Biochemistry; Superoxide Dismutase; Supplementation; Testing; Thinking; Toxic effect; Yeasts; Zinc; anti aging; disulfide bond; gain of function; glutaredoxin; in vivo; mutant; oxidation; oxygen toxicity; polypeptide; prevent; programs; protective effect; tissue culture; tissue/cell culture; yeast genetics

DESCRIPTION (provided by applicant): Copper and zinc containing superoxide dismutase (SOD1) plays a critical role in anti-oxidant defense, yet mutations in SOD1 can lead to amyotrophic lateral sclerosis (ALS), or Lou Gehrigs disease, through a toxic gain of function. The underlying mechanism remains unclear, but an emerging theme involves SOD1 misfolding and formation of toxic SOD1 aggregates. A hypothesis of this research is that the fate of ALS mutant SOD1 is governed by intracellular factors that control post-translational modification of the polypeptide. We have identified a number of such factors, including the CCS copper chaperone, a CCS-independent pathway for activating SOD1, and cytosolic glutaredoxins that help reduce the SOD1 disulfide. Precisely how these factors impact on SOD1 in vivo will be addressed in two Aims of the current proposal: Aim1-To understand the pathways for activating human SOD1 with copper: A combination of genetics, biochemistry and structural analyses will explore how human SOD1 discerns between two pathways for acquiring copper and how CCS may uniquely promote SOD1 folding through proline isomerization. Aim2-To understand the impact of SOD1 interacting factors on the stability and aggregation of SOD1: Molecular genetic studies in yeast and tissue culture will dissect the opposing roles of the copper activation pathways versus the disulfide reduction pathways in the folding and aggregation of ALS mutant SOD1. In a separate but related line of research, this proposal will also address an unprecedented role for CCS in chromatin silencing: Aim 3-To understand the dual roles of CCS in SOD1 activation and chromatin silencing. Yeast genetics and biochemistry will address how CCS can promote both SOD1 activation and chromatin silencing through a possible mechanism involving the anti-aging factor SIR2. Together, these studies in both yeast and mammalian cells promise to shed new light into the biology of SOD1 and its metallochaperone CCS with important implications to disease.

描述（由申请人提供）：含铜和锌的超氧化物歧化酶 (SOD1) 在抗氧化防御中发挥着关键作用，但 SOD1 的突变可通过毒性功能获得导致肌萎缩侧索硬化症 (ALS) 或 Lou Gehrigs 病。潜在的机制尚不清楚，但一个新兴的主题涉及 SOD1 错误折叠和有毒 SOD1 聚集体的形成。这项研究的一个假设是，ALS 突变体 SOD1 的命运是由控制多肽翻译后修饰的细胞内因子控制的。我们已经确定了许多这样的因素，包括 CCS 铜伴侣、独立于 CCS 的激活 SOD1 的途径，以及有助于减少 SOD1 二硫化物的胞质谷氧还蛋白。这些因素在体内对 SOD1 的准确影响将在当前提案的两个目标中得到解决： 目标 1-了解用铜激活人类 SOD1 的途径：遗传学、生物化学和结构分析的结合将探索人类 SOD1 如何辨别获取铜的两种途径以及 CCS 如何通过脯氨酸异构化独特地促进 SOD1 折叠。目标2-了解SOD1相互作用因素对SOD1稳定性和聚集的影响：酵母和组织培养中的分子遗传学研究将剖析铜活化途径与二硫键还原途径在ALS突变体SOD1折叠和聚集中的相反作用。 在一项单独但相关的研究中，该提案还将解决 CCS 在染色质沉默中前所未有的作用：目标 3 - 了解 CCS 在 SOD1 激活和染色质沉默中的双重作用。酵母遗传学和生物化学将解决 CCS 如何通过涉及抗衰老因子 SIR2 的可能机制促进 SOD1 激活和染色质沉默。 总之，这些在酵母和哺乳动物细胞中的研究有望为 SOD1 及其金属伴侣 CCS 的生物学提供新的线索，对疾病具有重要意义。