MOLECULAR ANALYSIS OF THE TRYPANOSOME GLYCOSOME

锥虫糖体的分子分析

• 批准号: 3133992
• 负责人: Marilyn Parsons
• 金额: $ 28.86万
• 依托单位: SEATTLE BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-12-01 至 1994-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3133992
• 关键词: Trypanosoma; antiprotozoal agents; biological signal transduction; computer graphics /printing; computer simulation; drug design /synthesis /production; gel electrophoresis; gene expression; genetic manipulation; genetic mapping; genetic regulation; glycolysis; hexokinase; high performance liquid chromatography; laboratory rabbit; laboratory rat; microorganism genetics; microorganism metabolism; molecular cloning; nucleic acid probes; nucleic acid sequence; organelles; parasitic disease chemotherapy; phosphoglycerate kinase; protozoal antigen; trypanosomiasis

Pathogenic African trypanosomes, the causative agent of African sleeping sickness, possess a unique subcellar organelle, the glycosome. This organelle, found only in members of the order Kinetoplastida (including the causative agents of Chagas disease and leishmaniasis) houses many of the enzymes of the Embden- Myerhof pathway of glycolysis. There is no vaccine against any of these diseases and chemotherapy is unfortunately toxic. A further understanding of the glycosomal organelle might lead to the development of targets for new chemotherapies. Glycosomal targeting signals and the regulation of glycosomal gene expression in Trypanosoma brucei will be studied. We have demonstrated translational control of expression of glycosomal phosphoglycerate kinase. The molecular mechanisms of this control will be analyzed. The work on gene regulation will be expanded to include a procyclic-specific glycosomal protein, malate dehydrogenase. The corresponding gene will be cloned, and the regulation of its expression investigated. Using an in vitro glycosomal protein import assay, we will characterize the signals on glycosomal proteins which are responsible for their proper localization in the cell. The requirements for signal function will be delineated both at the amino acid level and at the topographic level. By studying both phosphoglycerate kinase and malate dehydrogenase, hypotheses on the general nature of glycosomal targeting sequences may be formed and ultimately tested. The experiments proposed are designed to elucidate the molecular controls underlying the stage-specific phenotype and to provide an understanding of the properties of glycosomal targeting sequences. Insights into these crucial parameters of glycosomal biogenesis may identify unique aspects which may be selectively disrupted. This information would not only be useful in the specific case of Trypanosoma brucei, but may also provide clues for chemotherapy of diseases caused by Leishmania spp. and Trypanosoma cruzi.

致病性非洲锥虫，非洲锥虫病的病原体 昏睡病，拥有独特的亚细胞细胞器， 糖体。 这种细胞器仅存在于目成员中 动质体（包括恰加斯病的病原体 和利什曼病）含有许多 Embden 酶 Myerhof 糖酵解途径。 没有针对任何疾病的疫苗 不幸的是，这些疾病和化疗是有毒的。 一个 对糖体细胞器的进一步了解可能会导致 新化疗靶标的开发。 糖体 靶向信号和糖体基因的调控 将研究布氏锥虫中的表达。 我们有 证明了糖体表达的翻译控制 磷酸甘油酸激酶。 此现象的分子机制 控制进行分析。 基因调控方面的工作将是 扩展到包括前环特异性糖体蛋白， 苹果酸脱氢酶。 相应的基因将被克隆，并且 对其表达的调节进行了研究。 使用体外 糖体蛋白输入测定，我们将表征信号 糖体蛋白负责其正常 在细胞中的定位。 信号功能要求 将在氨基酸水平和 地形水平。 通过研究磷酸甘油酸激酶和 苹果酸脱氢酶，关于一般性质的假设 可以形成糖体靶向序列并最终 已测试。 所提出的实验旨在阐明 阶段特异性表型背后的分子控制 提供对糖体特性的理解 靶向序列。 深入了解这些关键参数 糖体生物发生可能会识别独特的方面，这可能是 选择性地被破坏。 这些信息不仅有用 在布氏锥虫的具体情况下，但也可能提供 利什曼原虫引起的疾病化疗的线索。和 克氏锥虫。