Mitochondrial aconitase: Fe-S cluster biogenesis and interaction with mtDNA
线粒体顺乌头酸酶:Fe-S簇的生物发生及其与线粒体DNA的相互作用
基本信息
- 批准号:8044173
- 负责人:
- 金额:$ 30.45万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2008
- 资助国家:美国
- 起止时间:2008-04-01 至 2013-03-31
- 项目状态:已结题
- 来源:
- 关键词:ATP HydrolysisATP phosphohydrolaseAconitate HydrataseActive SitesAffectAgingApoproteinsBackBiogenesisCellsCitratesCitric Acid CycleDNA BindingDNA MaintenanceDataDefectEnzymesFerredoxinGenesGuanosine TriphosphateGuanosine Triphosphate PhosphohydrolasesHomologous GeneHumanHydrolysisIn VitroIronIsocitratesLifeMaintenanceMediatingMembraneMitochondriaMitochondrial DNAMitochondrial MatrixMolecularMolecular ChaperonesNADHNADPNuclearNucleotidesOligonucleotidesOxidation-ReductionOxidative StressOxidoreductasePathway interactionsPhosphotransferasesPhysiologyPlayProcessProductionProtein ImportProteinsRoleSaccharomyces cerevisiaeScaffolding ProteinSiteSourceSpecificityStagingSulfurTestingYeastscysteine desulfurasein vivoisocitratemutantoxidative damagepersulfidespublic health relevanceresearch studyyeast protein
项目摘要
DESCRIPTION (provided by applicant): Mitochondrial aconitase is highly sensitive to oxidative damage during aging. It reversibly converts citrate to isocitrate in the tricarboxylic acid (TCA) cycle. The cubane [4Fe-4S]2+ cluster of aconitase is essential for its catalytic activity, but it also renders the enzyme highly vulnerable to oxidative stress. We have discovered that isolated wild-type Saccharomyces cerevisiae mitochondria contain a nucleotide (GTP, NAD(P)H and ATP)- dependent machinery for iron-sulfur cluster (ISC) biogenesis of aconitase (Aco1p). The cluster biogenesis occurs in the mitochondrial matrix by a multi-step process requiring multiple components. In Aim I, we will biochemically dissect these steps and determine the nucleotide requirements of each. In Aims II-IV, we will study proteins that we found are directly relevant to nucleotide-dependent processes in ISC biogenesis of Aco1p. These include a GTPase (Mtg1p), a NADH kinase (Pos5p), a NADPH-requiring reductase (Arh1p), and an ATPase (Ssq1p). Mitochondria lacking any of these proteins are deficient in ISC biogenesis of Aco1p and display greatly reduced aconitase activity. However, the causal defects in these mutant mitochondria must be different since these proteins have different functions. Mtg1p is a GTPase in the mitochondrial matrix, and we will test our hypothesis that Mtg1p mediates the effects of matrix GTP on ISC biogenesis of Aco1p (Aim II). Pos5p is a NADH kinase and is required for NADPH production in the matrix. We will determine if the role of Pos5p in ISC biogenesis of Aco1p is mediated by its effects on NADPH levels in the matrix (Aim III). NADPH is likely utilized by reductase(s) such as ferredoxin reductase (Arh1p), which may provide reducing equivalents for one or more steps in cluster biogenesis, and this will be tested as part of Aim III. Ssq1p is an Hsp70 chaperone with ATPase activity in the matrix. Therefore some of the effects of ATP on ISC biogenesis of Aco1p are likely mediated by Ssq1p, and this will be explored (Aim IV). In addition to Ssq1p, yeast mitochondria contain an abundant Hsp70 (Ssc1p) that is involved in protein import. In contrast, human mitochondria contain a single Hsp70 (hSSC1), and thus it may participate in both protein import and ISC biogenesis, and we will test this possibility as part of Aim IV. Aco1p is essential for mitochondrial DNA (mtDNA) stability, and this activity is independent of its enzymatic activity in the TCA cycle. Aim V is to investigate interaction of yeast and human mitochondrial aconitases with mtDNA. We will determine if human mitochondrial aconitase, like yeast Aco1p, is also bifunctional and participates in the maintenance of mtDNA. Nucleotides and/or redox status of the matrix may play a critical role in distributing aconitase between the TCA cycle and mt-nucleoids, and we will examine these possibilities. The mechanism underlying the biogenesis of ISCs of mitochondrial aconitase in yeast is likely to be very similar to that in human. Homologs of most, if not all, yeast proteins that participate in the process also exist in human. Thus, conclusions from yeast studies proposed here will be informative to human physiology. Public Health Relevance: Mitochondrial aconitase requires a 4Fe-4S cluster for its enzymatic activity in the tricarboxylic acid (TCA) cycle and is highly sensitive to oxidative damage during aging. In yeast, aconitase is also essential for the maintenance of mitochondrial DNA, and this activity is independent of its catalytic activity in the TCA cycle. This proposal seeks to investigate the molecular mechanism of Fe-S cluster biogenesis of aconitase and its interaction with mitochondrial DNA.
描述(由申请人提供):线粒体刺激酶对衰老过程中的氧化损伤高度敏感。它可逆地将柠檬酸盐转化为三羧酸(TCA)循环中的异位酸盐。古巴[4FE-4S] 2+腺苷酶对其催化活性至关重要,但它也使该酶高度容易受到氧化应激的影响。我们已经发现,酿酒酵母分离的野生型糖含量含有核苷酸(GTP,NAD(P)H和ATP) - 依赖性的铁硫簇(ISC)丙OCONITAPE酶(ACO1P)的机械。簇生物发生发生在线粒体基质中,这是通过需要多个组件的多步骤过程。在目标I中,我们将在生化上剖析这些步骤,并确定每个步骤的核苷酸要求。在AIMS II-IV中,我们将研究我们发现的蛋白质与ACO1P的ISC生物发生中的核苷酸依赖性过程直接相关。这些包括GTPase(MTG1P),NADH激酶(POS5P),NADPH重新验还原酶(ARH1P)和ATPase(SSQ1P)。缺乏这些蛋白质中任何一种的线粒体缺乏ACO1P的ISC生物发生,并且显示出大大降低的刺刺酶活性。但是,由于这些蛋白质具有不同的功能,因此这些突变的线粒体中的因果缺陷必须有所不同。 MTG1P是线粒体基质中的GTPase,我们将测试MTG1P介导矩阵GTP对ACO1P ISC生物发生的影响的假设(AIM II)。 POS5P是NADH激酶,是基质中NADPH产生所必需的。我们将确定POS5P在ACO1P的ISC生物发生中的作用是否是由其对基质中NADPH水平的影响(AIM III)介导的。 NADPH可能是通过还原酶(例如铁蛋白还原酶(ARH1P))来利用的,这可能为聚类生物发生的一个或多个步骤提供还原的等效物,这将作为AIM III的一部分进行测试。 SSQ1P是矩阵中具有ATPase活性的HSP70伴侣。因此,ATP对ACO1P的ISC生物发生的某些影响很可能是由SSQ1P介导的,并且将探索这一点(AIM IV)。除SSQ1P外,酵母线粒体还含有涉及蛋白质进口的大量HSP70(SSC1P)。相比之下,人线粒体包含单个HSP70(HSSC1),因此它可能参与蛋白质导入和ISC生物发生,我们将作为AIM IV的一部分测试这种可能性。 ACO1P对于线粒体DNA(mtDNA)稳定性至关重要,并且该活性与其在TCA循环中的酶活性无关。目的V是研究酵母和人线粒体刺激酶与mtDNA的相互作用。我们将确定人线粒体刺激酶(如酵母ACO1P)是否也是双功能的,并且参与MTDNA的维持。基质的核苷酸和/或氧化还原状态可能在在TCA循环和MT核苷之间分配痤疮酶方面起关键作用,我们将检查这些可能性。酵母中线粒体刺激酶ISC的生物发生的基础机制可能与人类非常相似。大多数(如果不是全部)参与该过程的酵母蛋白的同源物也存在于人类中。因此,此处提出的酵母研究的结论将为人类生理学提供信息。 公共卫生相关性:线粒体刺激酶在三羧酸(TCA)周期中的酶活性需要4FE-4S簇,并且对衰老过程中的氧化损伤高度敏感。在酵母中,痤疮酶对于维持线粒体DNA也是必不可少的,并且该活性与TCA循环中其催化活性无关。该提案旨在研究a刺酶的Fe-S簇生物发生的分子机制及其与线粒体DNA的相互作用。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
DEBKUMAR PAIN其他文献
DEBKUMAR PAIN的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('DEBKUMAR PAIN', 18)}}的其他基金
Mitochondrial aconitase: Fe-S cluster biogenesis and interaction with mtDNA
线粒体顺乌头酸酶:Fe-S簇的生物发生及其与线粒体DNA的相互作用
- 批准号:
7466687 - 财政年份:2008
- 资助金额:
$ 30.45万 - 项目类别:
Mitochondrial aconitase: Fe-S cluster biogenesis and interaction with mtDNA
线粒体顺乌头酸酶:Fe-S簇的生物发生及其与线粒体DNA的相互作用
- 批准号:
7580968 - 财政年份:2008
- 资助金额:
$ 30.45万 - 项目类别:
Mitochondrial aconitase: Fe-S cluster biogenesis and interaction with mtDNA
线粒体顺乌头酸酶:Fe-S簇的生物发生及其与线粒体DNA的相互作用
- 批准号:
7797998 - 财政年份:2008
- 资助金额:
$ 30.45万 - 项目类别:
Mitochondrial aconitase: Fe-S cluster biogenesis and interaction with mtDNA
线粒体顺乌头酸酶:Fe-S簇的生物发生及其与线粒体DNA的相互作用
- 批准号:
8254428 - 财政年份:2008
- 资助金额:
$ 30.45万 - 项目类别:
Mitochondrial aconitase: Fe-S cluster biogenesis and interaction with mtDNA
线粒体顺乌头酸酶:Fe-S簇的生物发生及其与线粒体DNA的相互作用
- 批准号:
8707615 - 财政年份:2008
- 资助金额:
$ 30.45万 - 项目类别:
MGMLP DEPLETION ON MITOCHONDRIAL INNER MEMBRANE
线粒体内膜上的 MGMLP 耗尽
- 批准号:
6976427 - 财政年份:2004
- 资助金额:
$ 30.45万 - 项目类别:
相似海外基金
Mitochondrial aconitase: Fe-S cluster biogenesis and interaction with mtDNA
线粒体顺乌头酸酶:Fe-S簇的生物发生及其与线粒体DNA的相互作用
- 批准号:
7466687 - 财政年份:2008
- 资助金额:
$ 30.45万 - 项目类别:
Mitochondrial aconitase: Fe-S cluster biogenesis and interaction with mtDNA
线粒体顺乌头酸酶:Fe-S簇的生物发生及其与线粒体DNA的相互作用
- 批准号:
7580968 - 财政年份:2008
- 资助金额:
$ 30.45万 - 项目类别:
Mitochondrial aconitase: Fe-S cluster biogenesis and interaction with mtDNA
线粒体顺乌头酸酶:Fe-S簇的生物发生及其与线粒体DNA的相互作用
- 批准号:
7797998 - 财政年份:2008
- 资助金额:
$ 30.45万 - 项目类别:
Mitochondrial aconitase: Fe-S cluster biogenesis and interaction with mtDNA
线粒体顺乌头酸酶:Fe-S簇的生物发生及其与线粒体DNA的相互作用
- 批准号:
8254428 - 财政年份:2008
- 资助金额:
$ 30.45万 - 项目类别:
Structural Basis of Biological Membrane Protein Functions and Drug Resistance
生物膜蛋白功能和耐药性的结构基础
- 批准号:
10925999 - 财政年份:
- 资助金额:
$ 30.45万 - 项目类别: