MINING THE YEAST GENOME FOR ZINC REGULATED GENES

挖掘酵母基因组中的锌调控基因

• 批准号: 6385192
• 负责人: THOMAS J LYONS
• 金额: $ 3.24万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-06 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6385192
• 关键词: Saccharomyces cerevisiae; fungal genetics; genetic regulation; genetic regulatory element; metal metabolism; microarray technology; microorganism metabolism; transcription factor; zinc

Zinc is an essential micronutrient that is abundantly utilized inside eukaryotic cells, but despite the relative importance of zinc, very little is known about how eukaryotic organisms properly metabolize this ion. S. cerevisiae will be used as a model organism to study the genetics of zinc metabolism, since the entire genome of this organism has been sequenced. Also a zinc dependent transcriptional activator (ZAP1) has been identified in this organism that is responsible for regulating genes in zinc homeostasis. The purpose of this study will be to use genome wide analysis to define a ZAP1 regulon. To accomplish this task, DNA microarray technology will be used that allows for the determination of comparative expression levels of the entire yeast genome under different conditions, such as high and low zinc concentrations. Preliminary microarray experiments revealed that over 200 genes have significantly altered transcription levels under zinc deficiency. The general design of this proposal is to repeat and refine these studies to obtain more reliable data as well as to identify genes that are truly ZAP1 regulated rather than simply sensitive to the secondary effects of zinc deficiency. Once ZAP1 target genes have been identified several of the most interesting will be selected for further genetic study by independent conformation of the zinc responsiveness then by deletional analysis of said genes to determine their function.

锌是一种必需的微量营养素，在真核细胞内被大量利用，但尽管锌相对重要，但人们对真核生物如何正确代谢这种离子知之甚少。酿酒酵母将被用作研究锌代谢遗传学的模式生物，因为该生物的整个基因组已被测序。在这种生物体中还发现了一种锌依赖性转录激活因子（ZAP1），它负责调节锌稳态的基因。本研究的目的是利用全基因组分析来定义 ZAP1 调节子。为了完成这项任务，将使用 DNA 微阵列技术来确定整个酵母基因组在不同条件下（例如高和低锌浓度）的比较表达水平。初步的微阵列实验表明，在锌缺乏的情况下，超过 200 个基因的转录水平发生了显着改变。该提案的总体设计是重复和完善这些研究，以获得更可靠的数据，并确定真正受 ZAP1 调节的基因，而不是简单地对锌缺乏的副作用敏感。一旦ZAP1靶基因被鉴定出来，将通过锌反应性的独立构象选择几个最有趣的基因进行进一步的遗传研究，然后通过对所述基因的删除分析来确定它们的功能。