A quantitative imaging-based screen reveals the exocyst as a network hub connecting endocytosis and exocytosis.
A quantitative imaging-based screen reveals the exocyst as a network hub connecting endocytosis and exocytosis.
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DOI：
10.1091/mbc.e14-11-1527
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发表时间：
2015-07-01
影响因子：
3.3
通讯作者：
McCusker D
中科院分区：
文献类型：
--
作者：
Jose M;Tollis S;Nair D;Mitteau R;Velours C;Massoni-Laporte A;Royou A;Sibarita JB;McCusker D
内吞作用和胞吐作用的空间组织机制还不清楚。定量成像屏幕和高密度单囊泡跟踪用于识别突变体，是有缺陷的内吞和胞吐囊泡组织。该屏幕确定了外囊复合物在连接两个途径中的作用。内吞作用和胞吐作用的耦合是从受精到神经元活动和细胞极性的基本生物学过程的基础。然而，内吞作用和胞吐作用的空间组织的机制需要澄清。在芽殖酵母中使用基于定量成像的筛选，我们确定了89个突变体在一个或两个途径的定位中显示缺陷。高分辨率的单囊泡跟踪显示，内吞和外吞突变体she4和bud6改变后高尔基体囊泡动力学以相反的方式。内吞和外吞途径在极性建立期间显示出强的相互依赖性，而在极性维持期间则更加独立。系统分析确定了外囊复合物作为一个关键的网络枢纽，丰富的遗传相互作用的内吞和外吞组件。与对照细胞相比，外泌突变体显示出改变的内吞和高尔基体后囊泡动力学和散布的内吞和外泌结构域。这些数据是一致的一个重要作用，外泌在协调内吞和胞吐。
The mechanisms governing the spatial organization of endocytosis and exocytosis are ill defined. A quantitative imaging screen and high-density single-vesicle tracking are used to identify mutants that are defective in endocytic and exocytic vesicle organization. The screen identifies a role for the exocyst complex in connecting the two pathways. The coupling of endocytosis and exocytosis underlies fundamental biological processes ranging from fertilization to neuronal activity and cellular polarity. However, the mechanisms governing the spatial organization of endocytosis and exocytosis require clarification. Using a quantitative imaging-based screen in budding yeast, we identified 89 mutants displaying defects in the localization of either one or both pathways. High-resolution single-vesicle tracking revealed that the endocytic and exocytic mutants she4∆ and bud6∆ alter post-Golgi vesicle dynamics in opposite ways. The endocytic and exocytic pathways display strong interdependence during polarity establishment while being more independent during polarity maintenance. Systems analysis identified the exocyst complex as a key network hub, rich in genetic interactions with endocytic and exocytic components. Exocyst mutants displayed altered endocytic and post-Golgi vesicle dynamics and interspersed endocytic and exocytic domains compared with control cells. These data are consistent with an important role for the exocyst in coordinating endocytosis and exocytosis.