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Nutritional Copper Signaling and Homeostasis

营养铜信号传导和体内平衡

基本信息

批准号：
7988827
负责人：
SABEEHA MERCHANT
金额：
$ 5.94万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
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：
2009-12-17 至 2010-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7988827
关键词：
Abbreviations Active Sites Affinity Algae Animal Model Ankyrin Repeat Area Arts Assimilations Back Binding Binding Proteins Biochemical Biochemical Genetics C-terminal Cardiovascular Diseases Cell Fraction Cell membrane Cells Ceruloplasmin Chemistry Chlamydomonas Chlamydomonas reinhardtii Chromatography Co-Immunoprecipitations Copper Culture Media Cysteine Cytochromes DNA Binding DNA-Binding Proteins Dissection Electron Transport Enzymes Extracellular Matrix Family Gel Gene Expression Microarray Analysis Genetic Transcription Genome Genomics Germ Cells Growth Hemeproteins Hereditary Disease Homeostasis Human Hypoxia In Vitro Iron Location Malnutrition Matrix Metalloproteinases Menkes Kinky Hair Syndrome Metabolic Metals Methods Micronutrients Modeling Modification Molecular Genetics Monitor Mutagenesis Names Nuclear Export Nuclear Localization Signal Nutritional Oligonucleotide Microarrays Organism Oxidation-Reduction Oxygen Pathway interactions Pattern Peptide Hydrolases Phenotype Physiological Plastocyanin Proteins Proteomics RNA Interference Reaction Reactive Oxygen Species Regulation Research Research Personnel Response Elements Role Route Signal Pathway Signal Transduction Signal Transduction Pathway Site Sulfhydryl Compounds System Technology Testing Tetrapyrroles Trace Elements Zinc amine oxidase apoplastocyanin base cell type hypocupremia in vivo knock-down loss of function mutant novel nutrition permease promoter prototype response uptake yeast two hybrid system zygote

项目摘要

DESCRIPTION (provided by applicant): Copper is an essential micronutrient because of its role in enzymes that catalyze redox reactions. In humans, acquired copper-deficiency is rare because of the low daily requirement of copper, but copper-deficiency can accompany malnutrition, be associated with genetic disorders like Menkes syndrome or result from overconsumption of zinc supplements. And marginal copper deficiency, a contributing factor in cardiovascular disease, may not be uncommon. The long-term objective of this project is to i) discover the adaptive biochemical changes in copper-deficient cells in the context of specific cuproenzyme-containing pathways, ii) dissect the underlying regulatory mechanisms for achieving these changes, and iii) understand the physiological rationale for the adaptive modifications. Chlamydomonas reinhardtii is the model organism for the study because the well-defined synthetic growth medium facilitates studies of trace element homeostasis, and the full power of classical biochemical and genetic approaches can be combined with state of the art genomic and proteomic methods. The previous studies of copper-deficiency responses in Chlamydomonas established the concept of "back up" copper-independent enzymes that are expressed in -Cu cells to compensate for loss of function of cuproenzymes (e.g. cyt c6, as a back up for plastocyanin and Crd2 as a back up for the multicopper ferroxidase), and led to the discovery of Crrl, a novel DNA binding protein that is required for all known copper-deficiency responses and is involved also in maintaining copper homeostasis in a copper replete cell. In this project period, the investigators will: 1) identify CRD2, which is required for high affinity iron uptake in a copper-deficient cell, determine its biochemical function in relation to Fox1 (ferroxidase) and Ftr1 (iron permease) function, and monitor its expression and sub-cellular location in response to copper and iron nutrition; 2) dissect 3 domains in the copper regulator Crrl - the DNA-binding SBP domain, ankyrin repeats and the Cys-rich C-terminus - by mutagenesis and in vivo and in vitro functional analysis including CuRE binding, metal binding and redox state, and determine the pattern of copper-dependent Crrl regulation and location, to develop a model for its action as a regulator of copper nutrition and hypoxia; and 3) identify novel targets of copper-deficiency by specialized oligonucleotide microarray analyses of -Cu vs. +Cu cells and err I vs. wild-type cells, in parallel with sub-proteomic gel- and multi-dimensional chromatography-based analyses of copper-replete vs. copper-deficient cells.

描述（由申请人提供）：铜是一种必需的微量营养素，因为它在催化氧化还原反应的酶中起作用。在人类中，由于每日铜需求量较低，后天性铜缺乏症很少见，但铜缺乏症可能伴随营养不良，与门克斯综合征等遗传性疾病有关，或由过度食用锌补充剂引起。边缘性铜缺乏是心血管疾病的一个促成因素，可能并不罕见。该项目的长期目标是i）在特定的含铜酶途径的背景下发现铜缺乏细胞中的适应性生化变化，ii）剖析实现这些变化的潜在调控机制，以及iii）理解适应性修饰的生理学原理。莱茵衣藻是该研究的模式生物，因为定义明确的合成生长培养基有利于微量元素稳态的研究，并且经典生物化学和遗传方法的全部功能可以与最先进的基因组学和蛋白质组学方法相结合。以往对衣原体铜缺乏反应的研究建立了“备份”铜非依赖性酶的概念，这些酶在-Cu细胞中表达以补偿铜酶功能的丧失（例如cyt c6，作为质体蓝素的备份，Crd 2作为多铜铁氧化酶的备份），并导致Crrl的发现，一种新的DNA结合蛋白，是所有已知的铜缺乏反应所需的，也参与维持铜充足细胞中的铜稳态。在本项目期间，研究者将：1）鉴定CRD 2，其是在铜缺乏细胞中高亲和力铁摄取所必需的，确定其与Fox 1相关的生化功能（铁氧化酶）和Ftr 1（铁通透酶）功能，并监测其响应铜和铁营养的表达和亚细胞定位; 2）通过诱变和包括CuRE结合、金属结合和氧化还原状态在内的体内和体外功能分析，解剖铜调节剂Crrl中的3个结构域-DNA结合SBP结构域、锚蛋白重复序列和富含Cys的C末端，并确定铜依赖性Crrl调节和定位的模式，以开发其作为铜营养和缺氧调节剂的作用模型;和3）通过-Cu与+Cu细胞和err I与野生型细胞的专门寡核苷酸微阵列分析鉴定铜缺乏的新靶点，与基于亚蛋白质组学凝胶和多维色谱的分析的铜-充足与铜-缺乏细胞平行。

项目成果

期刊论文数量（54）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Dissecting the contributions of GC content and codon usage to gene expression in the model alga Chlamydomonas reinhardtii.

DOI：
10.1111/tpj.13033
发表时间：
2015-11
期刊：
The Plant journal : for cell and molecular biology
影响因子：
0
作者：
Barahimipour R;Strenkert D;Neupert J;Schroda M;Merchant SS;Bock R
通讯作者：
Bock R

The Chlamydomonas genome reveals the evolution of key animal and plant functions

DOI：
10.1126/science.1143609
发表时间：
2007-10-12
期刊：
SCIENCE
影响因子：
56.9
作者：
Merchant, Sabeeha S.;Prochnik, Simon E.;Grossman, Arthur R.
通讯作者：
Grossman, Arthur R.

A revised mineral nutrient supplement increases biomass and growth rate in Chlamydomonas reinhardtii.

DOI：
10.1111/j.1365-313x.2011.04537.x
发表时间：
2011-06
期刊：
The Plant journal : for cell and molecular biology
影响因子：
0
作者：
Kropat J;Hong-Hermesdorf A;Casero D;Ent P;Castruita M;Pellegrini M;Merchant SS;Malasarn D
通讯作者：
Malasarn D

A subset of the diverse COG0523 family of putative metal chaperones is linked to zinc homeostasis in all kingdoms of life.