Frascati-mediated Iron Metabolism in Erythroblasts

这项建议的目的是确定相互作用的蛋白质伙伴和生化途径 Frascati线粒体金属转运蛋白及其在红系发育中的作用。生物化学和 参与发育中的红细胞线粒体铁代谢的蛋白质知之甚少。 斑马鱼Frascati基因突变导致严重的低色素性贫血和成熟停滞 促红细胞阶段。使用位置克隆策略，我们发现Frascati编码了一部小说 线粒体溶质载体家族的成员(SLC25)。Frascati基因在胎儿和胎儿组织中高度表达 斑马鱼和小鼠的成体造血组织。由Frascati缺失的ES细胞产生的红细胞 表现出成熟停滞，55Fe在血红素中的结合严重受损。小鼠产生突变 通过有针对性地破坏Frascati基因座死于严重贫血，从而证实其在 哺乳动物的红系发育。我们发现Frascati是把铁输送到 血红素和铁-S的线粒体聚集在发育中的红系细胞中的生物合成。这样做的目的是 建议确定Frascati转运蛋白的生化途径和相互作用的蛋白质伙伴 进一步加深我们对红细胞发育过程中线粒体铁代谢的理解。身份的鉴定 Frascati作为一种重要的线粒体铁转运蛋白及其相互作用蛋白将为 探索人类铁代谢和红细胞生成障碍的遗传工具。我们建议实现 这些目标有以下两个具体目标： 具体目标1：将确定Frascati转运蛋白的蛋白质相互作用伙伴并对其进行表征。 特定目标2：斑马鱼胚胎和小鼠细胞的比较转录组分析 弗拉斯卡蒂运输机将进行。

The objective of this proposal is to identify the interacting protein partners and biochemical pathway of the frascati mitochondrial metal transporter and their role in erythroid development. The biochemistry and proteins involved in mitochondrial iron metabolism of developing erythroblasts are poorly understood. Mutations in the zebrafish frascati gene lead to a profound hypochromic anemia and maturation arrest at the pro-erythroblast stage. Using a positional cloning strategy, we showed that the frascati encodes a novel member of the mitochondrial solute carrier family (SLC25). The frascati gene is highly expressed in fetal and adult hematopoietic tissues of zebrafish and mouse. Erythroblasts generated from frascati-null ES cells exhibit maturation arrest with severely impaired incorporation of 55Fe into heme. Mouse mutants generated by targeted disruption of the frascati locus die of severe anemia, thereby confirming its essential role in mammalian erythroid development. We showed that frascati is the major transporter for delivering iron into the mitochondria for heme and Fe-S cluster biosynthesis in developing erythroid cells. The goal of this proposal is to identify the biochemical pathway and interacting protein partners of the frascati transporter to further our understanding of mitochondrial iron metabolism in developing red cells. The identification of frascati as an essential mitochondrial iron transporter and its interacting proteins will provide powerful genetic tools for exploring human disorders of iron metabolism and erythropoiesis. We propose to achieve these aims by the following two specific aims: Specific Aim 1: Protein interacting partners of the frascati transporter will be identified and characterized. Specific Aim 2: A comparative transcriptome analysis of zebrafish embryos and mouse cells deficient for the frascati transporter will be conducted.