Ayala Inmaculada, Colanzi Antonino
Institute of Protein Biochemistry, National Research Council of Italy, Via P. Castellino 111, 80131, Naples, Italy.
Methods Mol Biol. 2016;1496:173-85. doi: 10.1007/978-1-4939-6463-5_14.
The Golgi complex of mammalian cells is composed of stacks of flattened cisternae that are connected by tubules to form a continuous membrane system, also known as the Golgi ribbon. At the onset of mitosis, the Golgi ribbon is progressively fragmented into small tubular-vesicular clusters and it is reconstituted before completion of cytokinesis. The investigation of the mechanisms behind this reversible cycle of disassembly and reassembly has led to the identification of structural Golgi proteins and regulators. Moreover, these studies allowed to discover that disassembly of the ribbon is necessary for cell entry into mitosis. Here, we describe an in vitro assay that reproduces the mitotic Golgi fragmentation and that has been successfully employed to identify many important mechanisms and proteins involved in the mitotic Golgi reorganization.
哺乳动物细胞的高尔基体复合体由扁平的囊泡堆叠组成，这些囊泡通过小管相连，形成一个连续的膜系统，也称为高尔基体带。在有丝分裂开始时，高尔基体带逐渐破碎成小的管状囊泡簇，并在胞质分裂完成前重新组装。对这种可逆的拆解和重新组装循环背后机制的研究，已经鉴定出了高尔基体结构蛋白和调节因子。此外，这些研究还发现，高尔基体带的拆解是细胞进入有丝分裂所必需的。在这里，我们描述了一种体外测定方法，该方法可重现有丝分裂期高尔基体的碎片化过程，并已成功用于鉴定许多参与有丝分裂期高尔基体重组的重要机制和蛋白质。
