Eukaryotic Heme Utilization

真核血红素利用

• 批准号: 7967561
• 负责人: Caroline Philpott
• 金额: $ 30.13万
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7967561
• 关键词: Ataxia; Binding; Biochemical; Biological; Caenorhabditis elegans; Cell membrane; Cells; Collaborations; Copper; Enzymes; Eukaryota; Exhibits; Genes; Genetic; Genetic Screening; Heme; Heme Iron; Hemoglobin; Hereditary Disease; Hereditary hemochromatosis; Homeostasis; Human; Hypoxia; Intracellular Transport; Iron; Iron Compounds; Iron Overload; Mammals; Maryland; Metabolism; Microarray Analysis; Myoglobin; Nutrient; Organic Iron Compounds; Organism; Oxygen; Porphyrins; Process; Regulation; Role; Saccharomyces cerevisiae; Saccharomycetales; System; Toxin; Universities; Yeasts; Zinc; cofactor; deprivation; design; heme a; human disease; insight; iron metabolism; loss of function; metal metabolism; microorganism; mutant; protoporphyrin IX; response; trafficking; uptake; yeast protein

Very little is known about heme transport in eukaryotes and a heme-specific transport uptake systems have not been identified in yeast. Although S. cerevisiae does not take up heme in response to iron deprivation, we have determined that this species does have an inducible heme uptake system that is activated under heme deficiency and hypoxic conditions. We performed microarray analysis of strains grown under these conditions, and found that a number of putative transporters are transcriptionally activated. We have identified transporters that, when over expressed, increase heme uptake into yeast. We have also identified a transporter, termed PUG1 (Porphyrin uptake gene 1), that stimulates the uptake of protoporphyrin IX, the immediate precursor of heme, and simultaneously inhibits the utilization of heme. This transporter is transcriptionally activated by oxygen and heme deficiency and is expressed on the plasma membrane of yeast. We have designed a genetic screen for the identification of yeast proteins involved in intracellular heme trafficking and are evaluating candidates. In collaboration with Iqbal Hamza of the University of Maryland, we are functionally characterizing the putative heme transporters of C. elegans by expressing them in yeast. Heme transporters from C. elegans have heme uptake activity when expressed in yeast and mutant forms of heme transporters exhibit loss of function when expressed in yeast.

关于血红素在真核生物中的转运知之甚少，并且在酵母中还没有鉴定出血红素特异性转运摄取系统。虽然S.虽然酿酒酵母对铁缺乏没有反应，但我们已经确定该物种确实具有在血红素缺乏和缺氧条件下激活的诱导型血红素摄取系统。我们对在这些条件下生长的菌株进行了微阵列分析，发现许多推定的转运蛋白被转录激活。我们已经确定了转运蛋白，当过度表达，增加血红素摄取到酵母。我们还确定了一种转运蛋白，称为PUG 1（卟啉摄取基因1），它刺激血红素的直接前体原卟啉IX的摄取，同时抑制血红素的利用。这种转运蛋白在缺氧和血红素缺乏时被转录激活，并在酵母的质膜上表达。我们已经设计了一个遗传筛选，用于鉴定参与细胞内血红素运输的酵母蛋白，并正在评估候选人。 我们与马里兰州大学的伊克巴尔·哈姆扎合作，对C.通过在酵母中表达它们。血红素转运蛋白C.当在酵母中表达时，秀丽线虫具有血红素摄取活性，并且当在酵母中表达时，血红素转运蛋白的突变形式表现出功能丧失。