RUI: Regulation Of Pyrimidine Metabolism In Plants

RUI：植物中嘧啶代谢的调节

• 批准号: 0516544
• 负责人: Robert Slocum
• 金额: --
• 依托单位: Goucher College
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0516544&HistoricalAwards=false
• 关键词: RUI; Regulation; Pyrimidine; Metabolism; Plants

Pyrimidine nucleotides are precursors in the biosynthesis of DNA and RNA, molecules which contain the essential genetic information for growth and development in plants and all other living organisms. UDP-sugars are also activated intermediates in the synthesis of certain types of lipids, as well as a major carbohydrate storage product (sucrose) and cell wall polysaccharides (cellulose, hemicellulose, pectins) in plants, accounting for much of the earth's biomass and production of food and fiber. Biochemical studies have helped to define major enzymes and pathways comprising pyrimidine metabolism, but plants remain the only major group of organisms for which basic mechanisms controlling pyrimidine metabolism have not been elucidated.A functional genomics approach will be used to study pyrimidine metabolism in the model plant, Arabidopsis. Using bioinformatics tools to "mine" information from integrated genomics databases, 146 genes encoding functionally characterized or putative enzymes in each of the major areas of pyrimidine metabolism (de novo synthesis of nucleotides, nucleotide interconversions, salvaging of preformed nucleobases/nucleosides to nucleotides, catabolic pathways) and related pathways (UDP-glucose and arginine synthesis) have been identified. The goals of this research are to 1) use DNA microarrays to characterize coordination of pathway activities, at the level of gene expression, and to identify potentially-regulated genes 2) to functionally characterize a small number of genes/enzymes in vitro (complementation of defined mutants, biochemical studies) or in planta (T-DNA knockouts, RNAi-mediated gene silencing, 35S promoter-driven overexpression in transgenic Arabidopsis lines) 3) to study expression of each gene in response to changes in pyrimidine availability in plant tissues (N-(phosphonacetyl)-L-aspartate (PALA) inhibition of de novo synthesis, exogenous pyrimidines, etc.) The proposed collaboration with The Institute for Genomic Research ensures that high-quality array data will be generated, while availability of full-length cDNAs/ESTs and Arabidopsis T-DNA knockout mutants for the enzymes to be studied will facilitate the proposed research.Broader impacts: These studies will significantly advance our understanding of the regulation of pyrimidine metabolism in plants and will provide important insights into coordination of these activities with other areas of plant metabolism. The project will provide training opportunities for undergraduate students in bioinformatics and plant functional genomics. Gene expression data from array experiments will also be used in future student investigator-driven projects in a bioinformatics course at Goucher College, expanding the educational impact of this research. Students will present their research at scientific meetings and will co-author publications. If the past trends continue, most of them will pursue post-baccalaureate training at top-ranked graduate or professional schools.

嘧啶核苷酸是DNA和RNA生物合成的前体，DNA和RNA分子包含植物和所有其他生物体生长和发育所必需的遗传信息。UDP-糖也是某些类型脂质合成中的活化中间体，以及植物中主要的碳水化合物储存产物（蔗糖）和细胞壁多糖（纤维素、半纤维素、果胶），占地球生物质和食物和纤维生产的大部分。生物化学的研究已经帮助确定了嘧啶代谢的主要酶和途径，但植物仍然是唯一一个尚未阐明嘧啶代谢基本机制的主要生物类群。使用生物信息学工具从综合基因组学数据库中“挖掘”信息，已经鉴定了146个基因，其编码在嘧啶代谢的每个主要领域（核苷酸的从头合成、核苷酸互变、将预先形成的核碱基/核苷抢救为核苷酸、分解代谢途径）和相关途径（UDP-葡萄糖和精氨酸合成）中的功能表征的或推定的酶。本研究的目的是1）使用DNA微阵列来表征基因表达水平上的通路活动的协调，并识别潜在的调控基因2）在体外功能上表征少量基因/酶（确定突变体的互补，生物化学研究）或在植物中（T-DNA敲除，RNAi介导的基因沉默，转基因拟南芥系中35 S启动子驱动的过表达）3）研究每种基因响应于植物组织中嘧啶可用性变化的表达（N-（膦酰基乙酰基）-L-天冬氨酸（PALA）抑制从头合成，外源性嘧啶等）拟议中的与基因组研究所的合作确保了高质量的阵列数据将被生成，而用于待研究酶的全长cDNA/EST和拟南芥T-DNA敲除突变体的可用性将促进拟议中的研究。这些研究将显著地推进我们对植物中嘧啶代谢调节的理解，并将为这些调节的协调提供重要的见解。植物代谢的其他领域。该项目将为本科生提供生物信息学和植物功能基因组学方面的培训机会。来自阵列实验的基因表达数据也将用于Goucher学院生物信息学课程中未来的学生演示器驱动项目，扩大这项研究的教育影响。学生将在科学会议上展示他们的研究，并将共同撰写出版物。如果过去的趋势继续下去，他们中的大多数人将在一流的研究生院或专业学校接受学士后培训。