Golgi Architecture, Biogenesis and Function

高尔基体结构、生物起源和功能

• 批准号: 7013096
• 负责人: GRAHAM B WARREN
• 金额: $ 39.91万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-01-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7013096
• 关键词: Golgi apparatus; SDS polyacrylamide gel electrophoresis; Toxoplasma gondii; Trypanosoma brucei; cell cycle; electron microscopy; fluorescence microscopy; immunoelectron microscopy; laboratory rat; mass spectrometry; molecular stacking; phosphorylation; polymerase chain reaction; protein binding; protein kinase; protein reconstitution; protein structure function; protein transport; regulatory gene; synthetic peptide; tissue /cell culture; vesicle /vacuole

DESCRIPTION (provided by applicant): The biogenesis of organelles ensures propagation through successive generations but our knowledge of the underlying mechanisms is patchy, especially for membrane-bound organelles such as the Golgi apparatus. There are two popular models. The first considers the Golgi to be an autonomous organelle, responsible for its own duplication and partitioning during mitosis. This is the model we favor. The second model considers the Golgi to be a dependent organelle, in dynamic steady state with the ER, and reliant on it for all aspects of biogenesis. In the last granting period we studied those proteins that tether vesicles to Golgi membranes and cisternal membranes to each other. Collectively, we termed these matrix proteins and were able to provide evidence that they constituted an underlying scaffold that is normally populated by enzyme containing membranes. In this renewal we aim to confirm and extend these observations, focusing on the GRASP family of stacking proteins. Our aim is to determine precisely how these proteins carry out their stacking function. We also want to understand the process of Golgi assembly, both at the end of mitosis when the partitioned Golgi is reassembled, and during subsequent duplication, when a new copy of the Golgi is made. In particular we want to know whether the process is "seeded" by matrix proteins and whether there is a precise order of cisternal assembly. Lastly, we have begun to study the process of Golgi duplication in two protozoan parasites, Toxoplasma gondii, and Trypanosoma brucei. Both have a single Golgi apparatus, the duplication of which can be followed in real time using video fluorescence microscopy. These simple systems are also medically important.

描述（由申请人提供）：细胞器的生物发生确保了连续的世代传播，但是我们对基本机制的了解是斑驳的，尤其是对于膜结合的细胞器，例如高尔基体设备。有两个流行的型号。第一个认为高尔基是一个自主细胞器，负责其在有丝分裂过程中进行重复和分区。这是我们喜欢的模型。第二个模型将高尔基人视为一个依赖的细胞器，在动态稳态下具有ER，并依赖于生物发生的各个方面。在最后的给予期间，我们研究了那些将囊泡绑在高尔基膜和脑膜膜上的蛋白质。总的来说，我们称这些基质蛋白质，并能够提供证据表明它们构成了通常由含膜酶填充的基础支架。在这种续签中，我们旨在确认和扩展这些观察结果，重点关注堆叠蛋白的掌握家族。我们的目的是精确确定这些蛋白质如何执行其堆叠功能。我们还希望了解高尔基体组装的过程，无论是在重新组装分区的高尔基体时，都在有丝分裂结束时，以及随后的重复，当制作了高尔基的新副本时。特别是我们想知道该过程是否是由基质蛋白“播种”的，以及是否存在精确的蓄水组组装顺序。最后，我们已经开始研究两个原生动物寄生虫，弓形虫弓形虫和布鲁氏锥虫的高尔基复制过程。两者都有一个高尔基体，可以使用视频荧光显微镜实时遵循其重复。这些简单的系统在医学上也很重要。