Mechanism of the Transcriptional Response to Transition Metals

过渡金属的转录反应机制

基本信息

批准号：
8269978
负责人：
MICHAEL Thomas MARR
金额：
$ 30.53万
依托单位：
BRANDEIS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-01 至 2014-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8269978
关键词：
Address Affect Binding Binding Proteins Binding Sites Biological Biological Models Cadmium Cd thionein Cell physiology Cells Complex Connective Tissue Copper Discrimination Disease Dose Drosophila genome Drosophila genus Elongation Factor Equilibrium Free Radicals Gene Expression Profile Gene Expression Regulation Gene Targeting Generations Genes Genetic Transcription Genomics Goals Growth Health Hepatolenticular Degeneration Homeostasis Human Immunoprecipitation Individual Iron Kidney Knowledge Lead Left Libraries Liver Mammalian Cell Mediator of activation protein Menkes Kinky Hair Syndrome Mercury Metabolic Metallothionein Metals Microarray Analysis Modeling Nature Nervous system structure Normal Cell Nutrient Nutritional Requirements Organism Pathology Physiological Play Positioning Attribute Proteins RNA Interference Regulation Regulon Response Elements Role Sequence-Specific DNA Binding Protein Signal Transduction Staging Stimulus Stress System Toxic effect Transcript Transcription Elongation Transcription Initiation Transcriptional Activation Transcriptional Regulation Transition Elements Work Zinc base brain tissue cohort in vitro Assay interest prevent programs promoter response sensor toxic metal transcription factor transcription factor MTF-1

项目摘要

Essential divalent transition metals such as zinc, copper, and iron play important structural and catalytic roles in over 300 proteins. However, these and other transition metals also pose a potential threat to an organism. Left unchecked these metals can catalyze the generation of free radicals that damage all types of biological molecules. Transition metals must be kept in a physiological window. Diseases like Wilson's disease and Menke's disease demonstrate that either too much or too little metal can lead to pathologies of the liver, kidney, nervous system and connective tissues. Clearly metal homeostasis is an important aspect of cellular function. A major part of this control occurs at the level of transcription. One of the central players in this regulation is the metal response element binding protein (MTF-1). MTF-1 is a sequence specific DNA binding protein that perceives the metal status of a cell and activates genes accordingly. A good deal is known about MTF-1 itself, but very little is known about the protein co-factors that help MTF-1 efficiently activate transcription. We will use an RNAi based screen, in the Drosophila model system, to determine what protein co-factors are required for metal stimulated transcription. In addition we will characterize the physical and functional interactions between MTF-1 and these protein co-factors using a combination of cell based and in vitro assays. Finally we will identify the MTF-1 binding sites across the Drosophila genome to define the MTF-1 regulon that responds to metal stimuli. The long-term objective of our studies is to understand how a cell, in response to a diverse set of metals, differentially regulates the appropriate metal responsive genes to control metal homeostasis.

必需的二价过渡金属，例如锌，铜和铁发挥重要的结构和催化在300多个蛋白质中的作用。但是，这些和其他过渡金属也对生物。剩下的这些金属可以催化损害所有类型的自由基的产生生物分子。过渡金属必须保存在生理窗口中。像威尔逊氏病这样的疾病 Menke的疾病表明，过多或太少的金属可以导致肝脏的病理，肾脏，神经系统和结缔组织。显然金属稳态是细胞的重要方面功能。该控制的主要部分发生在转录水平。其中的核心参与者之一调节是金属响应元件结合蛋白（MTF-1）。 MTF-1是序列特异性DNA结合感知细胞金属状态并相应地激活基因的蛋白质。一项众所周知 MTF-1本身，但对帮助MTF-1有效激活的蛋白质辅助因子知之甚少转录。我们将在果蝇模型系统中使用基于RNAi的屏幕来确定什么蛋白金属刺激的转录需要辅助因子。另外，我们将表征身体和 MTF-1与这些蛋白质辅助因子之间的功能相互作用，结合了基于细胞的组合和体外测定。最后，我们将确定跨果蝇基因组的MTF-1结合位点，以定义MTF-1 对金属刺激做出反应的法规。我们研究的长期目的是了解细胞如何响应对各种金属的响应，差异调节适当的金属响应基因以控制金属稳态。