The Biogenesis of the Golgi Apparatus in T.Brucei

描述（由申请人提供）： 高尔基体负责ER中生物合成的蛋白质和脂质的翻译后加工，然后将它们递送到细胞内外的不同位置。考虑到高尔基体在蛋白质运输中的重要性，其在细胞分裂期间的精确复制和遗传是精心策划的。在哺乳动物细胞中，高尔基体存在于一种更高级的结构中，称为高尔基带，这使得直接观察复制事件变得困难。简单的真核生物，包含一个或几个高尔基体堆栈，因此最近被用作模型系统来研究这一过程。沃伦实验室最近使用寄生虫布氏锥虫的工作提供了证据，证明旧的高尔基体提供了建立新细胞器所必需的成分，这表明其复制是一个模板化的过程。T. brucei将用于确定旧高尔基体和新内质网出口位点对构建新的功能性高尔基体的精确贡献。还将研究负责两个高尔基体之间运输的载体以及其他细胞器在定位新高尔基体中的作用。具体目标：（1）确定高尔基体基质蛋白是否在糖基转移酶之前转移到新高尔基体;（2）确定介导从旧高尔基体转移到新高尔基体的载体;（3）确定基体在新高尔基体遗传中的作用;（4）观察新高尔基体何时能够处理货物：（5）并确定在细胞周期晚期形成的“额外”高尔基体的作用。

DESCRIPTION (provided by applicant): The Golgi apparatus is responsible for post-translational processing of the proteins and lipids biosynthesized in the ER, and then delivering them to different locations within and outside the cell. Considering the importance of the Golgi in protein traffic, its accurate duplication and inheritance during cell division are carefully orchestrated. In mammalian cells the Golgi exists in a higher order structure that is known as the Golgi ribbon, making direct observation of the duplication event difficult. Simpler eukaryotes that contain one or just a few Golgi stacks have therefore been used recently as model systems to study this process. Recent work in the Warren lab using the parasite Trypanosoma brucei has provided evidence that the old Golgi provides components necessary for establishing the new organelle, suggesting that its duplication is a templated process. T. brucei will be used to determine the precise contributions of the old Golgi and the new ER exit site to the construction of a new, functional Golgi. The carriers responsible for transport between the two Golgi and the role of other organelles in positioning the new Golgi will also be investigated. Specific Aims: (1) To determine if Golgi matrix proteins are transferred to the new Golgi prior to a glycosyltransferase; (2) to identify the carriers that mediate transfer from the old Golgi to the new; (3) to identify the role of the basal body in the inheritance of the new Golgi; (4) to observe when the new Golgi becomes competent to process cargo: (5) and to determine the role of the 'extra' Golgi that form during the late cell cycle.