Genetic analysis of iron homeostasis in C. elegans

线虫铁稳态的遗传分析

基本信息

批准号：
7414595
负责人：
Elizabeth Ann Leibold
金额：
$ 20.41万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-05-15 至 2011-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7414595
关键词：
Affinity Animal Model Animals Be++ element Beryllium Binding Binding Sites Biochemical Biochemical Genetics Biological Process Caenorhabditis elegans Cells Chelating Agents Citric Acid Cycle Conserved Sequence Data Elements Enhancers Eukaryotic Cell Ferritin Fluorescence Free Radicals Gene Expression Gene Expression Profiling Genes Genetic Screening Genetic Transcription Goals Green Fluorescent Proteins Heme Homeostasis Homologous Gene Hybrids Iron Iron-Regulatory Proteins Learning Mediating Mutate Mutation Organ Organism Photosynthesis Proteins Regulation Regulatory Element Reporter Respiration Role Signal Pathway Signal Transduction System Tissues Transcriptional Regulation Transferrin Receptor Transgenes Yeasts base expectation genetic analysis interest macromolecule metabolic abnormality assessment mutant novel oxygen transport plant fungi promoter protein expression transcription factor uptake

项目摘要

DESCRIPTION (provided by applicant): Iron has an essential role in many biological processes, including DMA synthesis/respiration, the tricarboxylic acid cycle, oxygen transport, heme synthesis and photosynthesis. But iron can also be toxic due to its ability to generate free radicals that damage macromolecules in cells. Consequently, organisms have evolved precise mechanisms to sense, acquire and store iron. Our goal is to learn more about how eukaryotic cells sense iron, generate a signal and transduce the signal into regulating iron homeostasis. Specifically, we are interested in understanding how transcription factors sense iron, and regulate gene expression in animal cells. While iron-mediated transcription factors have been identified in fungi and plants, little is known about the mechanisms and the proteins regulating iron-mediated transcriptional regulation in animal cells. We have discovered that Caenorhabditis elegans express ferritin (CeFTN-1 and CeFTN-2), which are key iron storage proteins. Our data shows that Ceftn-1 and Ceftn-2 are transcriptionally regulated by iron, and that cis-regulatory sequences present in their 5' upstream regions are required for this regulation. We propose to identify the cis-regulatory sequences and the iron-sensing transcription factor that binds these elements, and to determine how this transcription factor senses iron. We will also identify other C. elegans genes involved in iron signaling pathways. C. elegans offers a complementary approach to iron metabolic studies in other organisms, in that they are amenable to genetic analysis, they have organ/tissue systems, and they have homologs of genes encoding proteins involved in mammalian iron homeostasis. The specific aims are 1) to characterize cis-acting iron-regulatory elements in Ceftn-1 and Ceftn-2 promoters, 2) to purify the C. elegans transcription factor that regulates the Ceftn-1 and Ceftn-2 genes, and to determine how this factor controls ferritin expression, and 3) to identify genes involved in iron signaling pathways in C. elegans using a green fluorescence protein (GFP)-based screen.

描述（由申请人提供）：铁在许多生物过程中具有重要作用，包括DMA合成/呼吸，三羧酸周期，氧转运，血红素合成和光合作用。但是铁也可能是有毒的，因为它能够产生自由基损害细胞中大分子的自由基。因此，生物已经发展出精确的机制，以感知，获取和存储铁。我们的目标是更多地了解真核细胞如何感知铁，产生信号并将信号转移到调节铁稳态中。具体而言，我们有兴趣了解转录因子如何感知铁和调节动物细胞中的基因表达。尽管在真菌和植物中已经鉴定出铁介导的转录因子，但对调节动物细胞中铁介导的转录调控的机制和蛋白质知之甚少。我们发现秀丽隐杆线虫表达铁蛋白（CEFTN-1和CEFTN-2），它们是关键的铁储存蛋白。我们的数据表明，CEFTN-1和CEFTN-2受铁的转录调节，并且该调节需要5'上游区域中存在的顺式调节序列。我们建议确定结合这些元素的顺式调节序列和铁感应转录因子，并确定该转录因子如何感觉到铁。我们还将确定参与铁信号通路的其他秀丽隐杆线虫基因。秀丽隐杆线虫为其他生物体的铁代谢研究提供了一种补充方法，因为它们可以接受遗传分析，它们具有器官/组织系统，并且具有编码参与哺乳动物铁稳态的蛋白质的基因同源物。 The specific aims are 1) to characterize cis-acting iron-regulatory elements in Ceftn-1 and Ceftn-2 promoters, 2) to purify the C. elegans transcription factor that regulates the Ceftn-1 and Ceftn-2 genes, and to determine how this factor controls ferritin expression, and 3) to identify genes involved in iron signaling pathways in C. elegans using a green fluorescence protein (GFP)-based screen.