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Structural Organization of the Actin Cytoskeleton at Sites of Clathrin-Mediated Endocytosis

机译:网格蛋白介导的内吞作用部位的肌动蛋白细胞骨架的结构组织。

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Background: The dynamic actin cytoskeleton plays an important role in clathrin-mediated endocytosis (CME). However, its exact functions remain uncertain as a result of a lack of high-resolution structural information regarding actin architecture at endocytic sites.Results: Using platinum replica electron microscopy in combination with electron tomography, we found that actin patches associated with clathrin-coated structures (CCSs) in cultured mouse cells consist of a densely branched actin network, in which actin filament barbed ends are oriented toward the CCS. The shape of the actin network varied from a small lateral patch at the periphery of shallow CCSs, to a collar-like arrangement around partly invaginated CCSs with actin filament barbed ends abutting the CCS neck, to a polarized comet tail in association with highly constricted or fully endocytosed CCSs.Conclusions: Our data suggest that the primary role of the actin cytoskeleton in CME is to constrict and elongate the bud neck and drive the endocytosed vesicles from the plasma membrane. Moreover, in these processes, barbed ends directly push onto the load, as in a conventional propulsion mechanism. Based on our findings, we propose a model for initiation, evolution, and function of the dendritic actin network at CCSs.
机译:背景:动态肌动蛋白细胞骨架在网格蛋白介导的内吞作用(CME)中起着重要作用。然而,由于缺乏有关内吞位点肌动蛋白结构的高分辨率结构信息,其确切功能仍然不确定。结果:使用铂复制电子显微镜结合电子断层扫描技术,我们发现肌动蛋白斑与网格蛋白涂层结构相关培养的小鼠细胞中的(CCS)由密集分支的肌动蛋白网络组成,其中肌动蛋白丝的倒钩端朝向CCS。肌动蛋白网络的形状从浅CCS周围的小侧向斑块到部分新生的CCS周围呈领状排列,肌动蛋白丝倒刺的末端紧靠CCS颈部,到极化的彗星尾巴伴有高度收缩或弯曲。结论:我们的数据表明肌动蛋白细胞骨架在CME中的主要作用是收缩和拉长了芽颈,并从质膜上驱动了内吞囊泡。此外,在这些过程中,与传统的推进机构一样,带刺的末端直接推到负载上。基于我们的发现,我们提出了CCSs的树突肌动蛋白网络的起始,进化和功能的模型。

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