ROLE OF THE ANTIZYME FAMILY DURING XENOPUS DEVELOPMENT

抗酶家族在非洲爪蟾发育过程中的作用

• 批准号: 7609960
• 负责人: Charles Richard Toth
• 金额: $ 2.38万
• 依托单位: UNIVERSITY OF RHODE ISLAND
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7609960
• 关键词: Anabolism; Binding; Catabolism; Cell Proliferation; Cells; Charge; Complement; Computer Retrieval of Information on Scientific Projects Database; Development; Disease; Enzymes; Family; Funding; Genes; Genetic Screening; Grant; Institution; Nature; Organism; Ornithine Decarboxylase; Pathway interactions; Phenotype; Play; Polyamines; Process; Proteins; Rate; Reaction; Regulation; Research; Research Personnel; Resources; Role; Saccharomyces cerevisiae; Saccharomycetales; Source; Toxic effect; United States National Institutes of Health; Xenopus; Yeasts; design; ornithine decarboxylase antizyme; uptake; yeast genetics

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Polyamines are charged organic molecules that are vitally important for macromolecular reactions in cells but are toxic at high levels. There exist well-regulated biosynthetic pathways for both polyamine anabolism and catabolism. A protein involved in the control of polyamine levels, antizyme (AZ), plays a role in regulating polyamine levels in cells. AZ lowers polyamines below toxic levels by inhibiting polyamine uptake and activity of the rate-limiting enzyme ornithine decarboxylase (ODC) via direct binding and targeted degradation of ODC. By understanding the function of AZ in a normal setting, one can better understand the role of AZ in a disease setting and determine the nature of its toxicity in controlling cell proliferation. A tractable organism for polyamine research is the budding yeast Saccharomyces cerevisiae. The regulation of polyamine synthesis in yeast has been delineated although not specifically the role of antizyme in the process. In addition, mammalian forms of the polyamine genes can complement their functional equivalents in yeast cells. Using a yeast genetic screen, genes that functionally interact with AZ and its regulation of ODC can be isolated and characterized. This study proposes to use a genetic screen to obtain yeast genes that play a role in the regulation and function of the ODC/AZ pathway in yeast. A strain has been designed that is dependent on ODC enzymatic activity for viability. Expression of antizyme results in induced lethality due to lowered levels of polyamines. We hope to isolate yeast genes that can interact with the polyamine pathway and revert the lethal phenotype.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 多胺是带电荷的有机分子，对于细胞中的大分子反应至关重要，但高浓度时具有毒性。 多胺合成代谢和分解代谢都存在调节良好的生物合成途径。 抗酶 (AZ) 是一种参与控制多胺水平的蛋白质，在调节细胞内多胺水平方面发挥着重要作用。 AZ 通过直接结合和定向降解 ODC，抑制多胺摄取和限速酶鸟氨酸脱羧酶 (ODC) 的活性，从而将多胺降低到毒性水平以下。 通过了解 AZ 在正常情况下的功能，人们可以更好地了解 AZ 在疾病情况下的作用，并确定其在控制细胞增殖方面的毒性性质。 用于多胺研究的易于处理的生物体是芽殖酵母酿酒酵母。 酵母中多胺合成的调节已经被描述，尽管没有具体描述抗酶在该过程中的作用。 此外，哺乳动物形式的多胺基因可以补充其在酵母细胞中的功能等同物。 使用酵母遗传筛选，可以分离和表征与 AZ 功能相互作用的基因及其对 ODC 的调节。 本研究拟通过基因筛选来获取在酵母中ODC/AZ途径的调控和功能中发挥作用的酵母基因。 已设计出一种依赖于 ODC 酶活性的菌株。 由于多胺水平降低，抗酶的表达导致诱导致死。 我们希望分离出可以与多胺途径相互作用并恢复致死表型的酵母基因。