权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Metal-Regulatory Factor 1 (MTF-1) Role in Development and Stress Response

金属调节因子 1 (MTF-1) 在发育和应激反应中的作用

基本信息

批准号：
8116531
负责人：
Matthew J. Jenny
金额：
$ 24.03万
依托单位：
UNIVERSITY OF ALABAMA IN TUSCALOOSA
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：
2009-09-03 至 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8116531
关键词：
2-tert-butylhydroquinone Binding Binding Proteins Bioinformatics Cadmium Cell Differentiation process Cellular Stress Chromatin Computer Analysis DNA Damage Development Dominant-Negative Mutation Dose Drug Metabolic Detoxication Embryo Embryonic Development Gene Combinations Gene Expression Gene Expression Profiling Gene Targeting Genes Germ Lines Heat-Shock Proteins 70 Homeostasis Hypoxia Investigation Ischemia Knock-out Larva Liver Measurement Metal exposure Metals Microarray Analysis Microinjections Modeling Molecular Profiling Mus Mutation Neurodegenerative Disorders Neurons Nuclear Oligonucleotide Microarrays Oligonucleotides Oxidative Stress Pancreas Phenotype Play Promoter Regions Property Protein Isoforms Proteins RNA Regulation Regulator Genes Repression Research Response Elements Reverse Transcriptase Polymerase Chain Reaction Role Signal Transduction Site Staging Stress Techniques Testing Time Tissue Differentiation Tissues Transactivation Transcriptional Activation Transcriptional Regulation Transgenic Organisms Zebrafish Zinc Zinc Fingers biological adaptation to stress enhanced green fluorescent protein functional genomics in vivo insight knock-down novel novel strategies prevent promoter relating to nervous system research study response stressor transcription factor tumor progression

项目摘要

The metal response element-binding transcription factor-1 (MTF-1) is a ubiquitously expressed transcription factor that plays a primary role in the regulation of a variety of genes involved in metal homeostasis, embryonic development, tumor progression, and oxidative stress or hypoxia signaling. The overall objective ofthis research is to characterize the overlapping role of MTF-1 with NRF2, the primary transcription factor for regulating the oxidative stress response, in development and transcriptional co-regulation of stress responsive genes. The specific aims ofthis project are to (1) determine role of MTF-1 in zebrafish embryonic development, (2) determine phenotypic changes in expression of known stress response genes and identify novel target genes via MTF-1 and NRF2 'knockdowns', and (3) determine the interactive role of MTF-1 in coregulating transcriptional activation of NRF2 target genes. A variety of techniques and approaches will be employed to achieve these aims. These techniques will include development of transgenic zebrafish lines, morpholino knockdown of gene expression, gene expression profiling with oligonucleotide microarrays, and advanced bioinformatic techniques for microarray analysis and computational promoter analysis. A novel transgenic zebrafish strain that expresses a constitutively nuclear, dominant-negative MTF-1 isoform and three transgenic zebrafish lines that express green or yellow fluorescent protein in the gut, liver, pancreas and neurons will be used to visualize potential developmental phenotypes associated with the MTF-1 'knockdowns'. Second, oligonucleotide microarrays will be used to determine changes in gene expression after MTF-1 or NRF2 'knockdown' to discover novel target genes co-regulated by both transcription factors during development or in response to cadmium or tBHQ (oxidative stress). Third, the interactive role of MTF-1 with NRF2 in the transcriptional response to metals or oxidative stress will be further characterized by selecting target genes for computational promoter analysis to confirm the presence of MTF-1 and NRF2 recognition sites, and one gene will be selected for additional characterization by in vivo promoter analysis in response to cadmium or oxidative stress.

金属反应元件结合转录因子-1 (MTF-1) 是一种普遍表达的转录因子，在调节涉及金属稳态、胚胎发育、肿瘤进展以及氧化应激或缺氧信号传导的多种基因中发挥主要作用。本研究的总体目标是表征 MTF-1 与 NRF2（调节氧化应激反应的主要转录因子）在应激反应基因的发育和转录共同调节中的重叠作用。该项目的具体目标是（1）确定MTF-1在斑马鱼胚胎发育中的作用，（2）确定已知应激反应基因表达的表型变化，并通过MTF-1和NRF2“敲低”鉴定新的靶基因，以及（3）确定MTF-1在共同调节NRF2靶基因转录激活中的相互作用作用。将采用各种技术和方法来实现这些目标。这些技术将包括转基因斑马鱼品系的开发、基因表达的吗啉代敲低、寡核苷酸微阵列的基因表达谱分析以及用于微阵列分析和计算启动子分析的先进生物信息学技术。一种表达组成型核显性失活 MTF-1 亚型的新型转基因斑马鱼品系和三种在肠道、肝脏、胰腺和神经元中表达绿色或黄色荧光蛋白的转基因斑马鱼品系将用于可视化与 MTF-1“敲低”相关的潜在发育表型。其次，寡核苷酸微阵列将用于确定 MTF-1 或 NRF2“敲低”后基因表达的变化，以发现在发育过程中或响应镉或 tBHQ（氧化应激）时由两种转录因子共同调节的新靶基因。第三，MTF-1与NRF2在对金属或氧化应激的转录反应中的相互作用作用将通过选择用于计算启动子分析的靶基因来进一步表征，以确认MTF-1和NRF2识别位点的存在，并且将选择一个基因通过响应镉或氧化应激的体内启动子分析进行额外表征。