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）：铜是必不可少的微量营养素，因为它在催化氧化还原反应的酶中的作用。在人类中，由于铜的每日需求较低，因此获得的铜缺陷很少见，但是铜缺陷可能伴随营养不良，与Menkes综合征（如Menkes综合征）或锌补充剂过度消耗导致的遗传疾病有关。边缘铜缺乏症是心血管疾病的促成因素，可能并不少见。该项目的长期目的是i）在特定的含杯状酶的途径的背景下发现铜缺陷细胞的适应性生化变化，ii）剖析实现这些变化的基本调节机制，而iiii）了解生理学。自适应修饰的理由。衣原体Reinhardtii是该研究的模型生物，因为明确定义的合成培养基促进了痕量元素稳态的研究，并且可以将经典生物化学和遗传方法的全部力量与ART基因组和蛋白质组学方法的状态相结合。关于衣原体中铜缺陷反应的先前研究，建立了在-cu细胞中表达的“备份”铜依赖性酶的概念，以补偿cyt c6的功能损失（例如，Cyt C6，作为质质蛋白和CRD2的备份作为多型铁氧化酶的备份），并导致发现CRRL，这是一种新型的DNA结合蛋白，是所有已知的铜缺陷响应所必需的，并且还参与了在铜细胞中维持铜稳态。在此项目期间，研究人员将：1）确定CRD2，这是铜缺陷细胞中高亲和铁吸收所必需的，确定其与FOX1（铁氧化酶）和FTR1（Iron Permence）功能相关的生化功能，并监视它的表达和亚细胞位置响应铜和铁营养； 2）通过诱变，体内和体外功能分析（包括固化结合，金属结合和redox态和redox态，以及氧化和氧化和氧化）状态分析，在铜调节剂CRRL中解剖3个结构域 - DNA结合SBP结构域，Ankyrin重复序列和富含Cys的C-末端确定依赖铜的CRRL调节和位置的模式，以开发一种模型，以作为其作为铜营养和缺氧的调节剂的作用； 3）通过与基于亚蛋白质组的凝胶和基于多维的色谱分析，通过专门的-CU与 +Cu细胞和ERR I与野生型细胞的专业寡核苷酸微阵列分析来确定铜缺陷的新目标。铜重复与缺陷铜细胞。

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

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

DOI：
10.1186/1471-2164-10-470
发表时间：
2009-10-12
期刊：
BMC genomics
影响因子：
4.4
作者：
Haas CE;Rodionov DA;Kropat J;Malasarn D;Merchant SS;de Crécy-Lagard V
通讯作者：
de Crécy-Lagard V

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

The ins and outs of algal metal transport.

DOI：
10.1016/j.bbamcr.2012.04.010
发表时间：
2012-09
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH
影响因子：
5.1
作者：
Blaby-Haas, Crysten E.;Merchant, Sabeeha S.
通讯作者：
Merchant, Sabeeha S.

Copper economy in Chlamydomonas: Prioritized allocation and reallocation of copper to respiration vs. photosynthesis

DOI：
10.1073/pnas.1422492112
发表时间：
2015-03-03
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
影响因子：
11.1
作者：
Kropat, Janette;Gallaher, Sean D.;Merchant, Sabeeha S.
通讯作者：
Merchant, Sabeeha S.