MOLECULAR ANALYSIS OF THE TRYPANOSOME GLYCOSOME

锥虫糖体的分子分析

• 批准号: 2390290
• 负责人: Marilyn Parsons
• 金额: $ 30.82万
• 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 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2390290
• 关键词: Trypanosoma; gel electrophoresis; gene expression; gene targeting; glycolysis; host organism interaction; intracellular parasitism; laboratory mouse; laboratory rat; leishmaniasis; microorganism genetics; microorganism metabolism; molecular cloning; nucleic acid probes; nucleic acid sequence; organelles; peroxisome; phosphoglycerate kinase; protein signal sequence; protein transport; trypanosomiasis

Members of the order Kinetoplastida, including the causative agents of African sleeping sickness (Trypanosoma brucei subspecies), leishmaniasis (Leishmania spp.), and Chagas disease (Trypanosoma cruzi) possess a unique subcellular organelle, the glycosome. This organelle houses many of the enzymes of the Embden-Meyerhof pathway of glycolysis, as wells as enzymes involved in nucleotide biosynthesis, ether-lipid biosynthesis, and carbon dioxide fixation. There is no vaccine against any of these diseases and chemotherapy is toxic. A further understanding of the glycosomal organelle might lead to the development of new chemotherapies, particularly in the case of T. brucei where the parasites rely completely on glycosomal metabolism for the generation of energy while in the mammalian host. Our studies are aimed at analyzing specific constituents of the glycosome and the glycosomal biogenesis system, to further elucidate unique aspects of glycosomal metabolism and its role in the survival of the organism. Using transient transfection and a luciferase reporter system, experiments will identify and analyze the signals on proteins which target them to the glycosome. Gene knockout and gene replacement studies will examine the role of a phosphoglycerate kinase found only in the glycosome of Kinetoplastida. Glycosome-deficient mutants of T. brucei and Leishmania will be generated and studied to determine if the lack of the glycosome attenuates the parasite. These studies will clarify the role of the glycosome in Leishmania, which do not rely as extensively on glycolysis as does T. brucei. In addition, the mutants will also serve as the starting point for genetic experiments aimed at isolating genes important in glycosomal biogenesis. Insight into these crucial parameters of glycosomal biogenesis may identify unique aspects which may be selectively disrupted, and will allow us to determine whether the glycosome is an appropriate target for chemotherapy of diseases caused by trypanosomes and Leishmania.

订单动力层的成员，包括 非洲睡眠（Brucei锥虫亚种），利什曼病 （Leishmania spp。）和Chagas病（Cruzi锥虫）具有独特的 亚细胞细胞器，糖体。 这个细胞器容纳了许多 糖酵解的胚胎杂种途径的酶，以及酶 参与核苷酸生物合成，醚 - 脂质生物合成和碳 二氧化碳固定。这些疾病中没有疫苗， 化疗是有毒的。 对糖体的进一步了解 Organelle可能导致新化学疗法的发展， 特别是在寄生虫完全依赖的T. Brucei的情况下 关于能量产生的糖体代谢 哺乳动物宿主。我们的研究旨在分析特定成分 糖体和糖体生物发生系统的 阐明糖体代谢的独特方面及其在 有机体的生存。使用瞬时转染和荧光素酶 记者系统，实验将识别和分析信号 将它们靶向糖体的蛋白质。基因敲除和基因 替代研究将检查磷酸甘油酸激酶的作用 仅存在于动质体的糖体中。糖体缺陷突变体 将生成并研究布氏锥虫和利什曼原虫的样本，以确定是否 糖体的缺乏会使寄生虫减弱。这些研究将 阐明糖体在利什曼原虫中的作用，这不依赖于 像布鲁氏菌一样广泛地对糖酵解。另外，突变体将 也作为基因实验的起点 分离糖体生物合成中重要的基因。洞悉这些 糖体生物发生的关键参数可能会识别独特的方面 这可能会被选择性地破坏，并使我们能够确定是否 糖体是化疗引起的疾病的合适靶点 由锥虫和利什曼尼亚。