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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：
7879781
负责人：
Matthew J. Jenny
金额：
$ 24.07万
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 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7879781
关键词：
2-tert-butylhydroquinone Actins Age Antisense RNA Attention Award Binding Bioinformatics Biological Assay Blast Cell C-terminal Cadmium Cell Differentiation process Cells Cellular Stress Classification Co-Immunoprecipitations Collaborations Coupled Cysteine DNA Binding DNA Damage Data Detection Development Developmental Delay Disorders Dominant-Negative Mutation Dose Doxycycline Drosophila genus Effectiveness Elements Embryo Embryonic Development Experimental Designs Figs - dietary Fishes Fluorescence Microscopy Gene Combinations Gene Expression Gene Expression Profiling Gene Expression Regulation Gene Targeting Generations Genes Genome Goals Heat-Shock Proteins 70 Homeostasis Hypoxia Imagery In Situ Hybridization Ischemia Knock-out Length Liver Long-Term Effects Maps Measurement Mediating Mentors Metallothionein Metals Methods Microarray Analysis Microinjections Molecular Profiling Mus Mutate NF-E2-related factor 2 Neurodegenerative Disorders Neurons Nuclear Nucleic Acid Regulatory Sequences Oligonucleotide Microarrays Ontology Operon Organogenesis Oxidation-Reduction Oxidative Stress Pancreas Pattern Phase Phenotype Play Protein Isoforms Proteins Publishing RNA Polymerase II Recombinants Regulation Regulator Genes Regulatory Element Reporter Reporter Genes Research Response Elements Reverse Transcriptase Polymerase Chain Reaction Role Sampling Scanning Signal Transduction Site Staging Stress System Techniques Technology Testing Tetanus Helper Peptide Tetracycline Resistance Tetracyclines Time Tissues Transactivation Transcript Transcriptional Activation Transcriptional Activation Domain Transgenic Mice Transgenic Organisms Xenopus Zebrafish Zinc Fingers advanced system analog base biological adaptation to stress in vivo indexing insight knock-down mortality novel nuclease promoter relating to nervous system research study response stressor success tool transcription factor transcriptomics transgene expression tumor progression zebrafish development zebrafish genome

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）是一种广泛表达的转录因子，其在涉及金属稳态、胚胎发育、肿瘤进展和氧化应激或缺氧信号传导的多种基因的调节中起主要作用。本研究的总体目标是表征MTF-1与NRF 2的重叠作用，NRF 2是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调节中。本项目的具体目标是（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NRF 2“敲低”鉴定新的靶基因，（3）确定MTF-1在共调节NRF 2靶基因转录激活中的相互作用。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系的发展，基因表达的吗啉敲低，寡核苷酸微阵列的基因表达谱，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。一种新的转基因斑马鱼品系，表达组成型核，显性负MTF-1亚型和三个转基因斑马鱼系，表达绿色或黄色荧光蛋白的肠道，肝脏，胰腺和神经元将被用来可视化与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NRF 2“敲低”后基因表达的变化，以发现在发育过程中或响应于镉或tBHQ（氧化应激）由两种转录因子共同调控的新靶基因。第三，MTF-1与NRF 2在对金属或氧化应激的转录应答中的相互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NRF 2识别位点的存在，并且将通过响应于镉或氧化应激的体内启动子分析来选择一个基因用于另外的表征。