Frascati: mitochondrial transporter and erythropoiesis

Frascati：线粒体转运蛋白和红细胞生成

• 批准号: 7211687
• 负责人: BARRY H PAW
• 金额: $ 35.88万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-01 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7211687
• 关键词: Adult; Biochemical; Defect; Development; Disease; Disruption; Embryo; Erythroblasts; Erythroid Cells; Erythropoiesis; Escherichia coli; Family; Gene Targeting; Genes; Goals; Human; Hypochromic anemia; Inclusion Bodies; Iron; Iron Overload; Iron deficiency anemia; Lead; Medical; Mitochondria; Mus; Mutation; Organism; Physiological; Property; Recombinant Proteins; Role; Staging; Testing; Zebrafish; fetal; hematopoietic tissue; immortalized cell; insight; iron metabolism; member; novel; positional cloning; proteoliposomes; reconstitution; solute

DESCRIPTION (provided by applicant): The objective of this proposal is to understand the role of frascati, a novel mitochondrial solute transporter (SLC25), in vertebrate erythropoiesis and iron metabolism. Acquired and congential defects in iron metabolism from either deficiency or excess are one of the most common medical diseases. Mutations in the zebrafish frascati gene lead to a profound hypochromic anemia and a developmental arrest at the pro- erythroblast stage. Using a positional cloning strategy, we showed that frascati encodes a member of the mitochondrial solute carrier family. The frascati gene (mitoferrin, slc25a37) is highly expressed in fetal and adult hematopoietic tissues of zebrafish and mouse. We hypothesize that frascati is responsible for delivering iron to the mitochondria, and in particular, the mitochondria of developing erythroid cells. The goal of this application is to test this hypothesis. We propose to accomplish this goal by the following two specific aims: Specific Aim 1: Analysis of the biochemical function of the frascati gene. 1.1 The role of the frascati gene in mitochondrial iron acquisition will be studied using immortalized cells and zebrafish embryos deficient in frascati activity. 1.2 The physiologic substrate and transport properties of the frascati mitochondrial transporter will be studied by expression of recombinant protein as inclusion bodies in E. coli and their reconstitution in proteoliposomes. Specific Aim 2: Analysis of the frascati gene in mammalian organisms. 2.1 The function of the frascati gene in mouse will be studied by targeted gene disruption. Insight into the function of the frascati gene will be directly relevant to our understanding of human disorders of iron deficiency anemia and iron-overload.

描述（由申请人提供）：本提案的目的是了解frascati，一种新型线粒体溶质转运蛋白（SLC25）在脊椎动物红细胞生成和铁代谢中的作用。后天和先天铁代谢缺陷，无论是缺铁还是缺铁，都是最常见的医学疾病之一。斑马鱼frascati基因的突变导致深度低色性贫血和红细胞前阶段的发育停止。利用位置克隆策略，我们发现frascati编码线粒体溶质载体家族的一个成员。frascati基因（丝裂铁蛋白，slc25a37）在斑马鱼和小鼠的胎儿和成年造血组织中高度表达。我们假设frascati负责将铁传递到线粒体，特别是发育中的红细胞的线粒体。这个应用程序的目标是测试这个假设。我们建议通过以下两个具体目标来实现这一目标：具体目标1：分析frascati基因的生化功能。1.1将使用永活细胞和缺乏frascati活性的斑马鱼胚胎研究frascati基因在线粒体铁获取中的作用。1.2将重组蛋白作为包涵体在大肠杆菌中表达，并在蛋白脂质体中重组，研究重组蛋白线粒体转运体的生理底物和转运特性。专项目标2：分析哺乳动物体内的frascati基因。2.1通过靶向基因破坏，研究frascati基因在小鼠体内的功能。深入了解frascati基因的功能将直接关系到我们对缺铁性贫血和铁超载等人类疾病的理解。