Golgi Architecture, Biogenesis and Function

描述（由申请人提供）：细胞器的生物发生确保了连续世代的繁殖，但我们对潜在机制的了解是不完整的，特别是对于膜结合的细胞器，如高尔基体。有两种流行的模式。第一种观点认为高尔基体是一个自主的细胞器，在有丝分裂过程中负责自身的复制和分配。这是我们喜欢的模式。第二个模型认为高尔基体是一个依赖的细胞器，在动态稳定状态与ER，并依赖于它的所有方面的生物发生。在最后一个授予期间，我们研究了那些蛋白质的束缚囊泡高尔基膜和池膜彼此。总的来说，我们将这些基质蛋白命名为基质蛋白，并能够提供证据表明它们构成了通常由含酶膜填充的基础支架。在这次更新中，我们的目标是确认和扩展这些观察结果，重点是GRASP家族的堆积蛋白。我们的目标是精确地确定这些蛋白质如何执行其堆积功能。我们还想了解高尔基体组装的过程，包括在有丝分裂结束时，被分割的高尔基体重新组装，以及在随后的复制过程中，当一个新的高尔基体被复制时。我们特别想知道这个过程是否是由基质蛋白“播种”的，以及是否有一个精确的脑池组装顺序。最后，我们已经开始研究两种原生动物寄生虫，弓形虫和布氏锥虫的高尔基体复制的过程。两者都有一个单一的高尔基体，其复制可以遵循真实的时间使用视频荧光显微镜。这些简单的系统在医学上也很重要。

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.