A dynamin-actin interaction is required for vesicle scission during endocytosis in yeast.
A dynamin-actin interaction is required for vesicle scission during endocytosis in yeast.
复制标题
DOI：
10.1016/j.cub.2015.01.061
复制
发表时间：
2015-03-30
期刊：
影响因子：
9.2
通讯作者：
Ayscough, Kathryn R.
中科院分区：
文献类型：
--
作者：
Palmer, Sarah E.;Smaczynska-de Rooij, Iwona I.;Marklew, Christopher J.;Allwood, Ellen G.;Mishra, Ritu;Johnson, Simeon;Goldberg, Martin W.;Ayscough, Kathryn R.
肌动蛋白对酵母细胞的内吞作用至关重要，在张力下的哺乳动物细胞也是如此。然而，肌动蛋白聚合产生的力如何传递到质膜以驱动内陷和断裂仍然存在问题。在这里，我们揭示了酵母动力蛋白Vps1结合和捆绑丝状肌动蛋白。对茎结构域螺旋上的Vps1的突变分析确定了一个突变体RR457-458EE，它与肌动蛋白的结合更弱。在体内对Vps1功能的分析表明，该突变破坏了Vps1的内吞作用，但不会破坏Vps1的其他功能，如空泡运输或过氧化物体分裂。突变体Vps1在细胞中稳定表达，并与内吞报告基因Abp1和两栖生物素Rvs167共定位。对个体内吞斑块行为的详细分析表明，该突变导致内吞作用后期的异常运动，与断裂缺陷一致。用电子显微镜对酵母细胞进行的超微结构分析显示，内陷深度显著增加，进一步支持了Vps1-肌动蛋白在分裂过程中的相互作用。在体外对突变蛋白的分析表明，像野生型Vps1一样，它能够形成寡聚环，但关键的是，它已经失去了将肌动蛋白细丝捆绑成高阶结构的能力。提出了一个模型，在该模型中，肌动蛋白细丝在内陷过程中与Vps1结合，这种相互作用对于将肌动蛋白聚合力传递到膜上以驱动成功断裂是重要的。酵母菌动力蛋白Vps1在体外形成与F-肌动蛋白结合和捆绑的寡聚环。在体内Palmer等人中，Vps1茎区域的突变破坏了肌动蛋白的结合，捆绑Vps1 RR-EE肌动蛋白结合突变体存在细胞内断裂缺陷。揭示酵母动力蛋白Vps1能够形成能够直接与F-肌动蛋白相互作用并结合的寡聚环。Vps1-肌动蛋白相互作用中的缺陷导致体内细胞内分裂缺陷。
Actin is critical for endocytosis in yeast cells, and also in mammalian cells under tension. However, questions remain as to how force generated through actin polymerization is transmitted to the plasma membrane to drive invagination and scission. Here, we reveal that the yeast dynamin Vps1 binds and bundles filamentous actin. Mutational analysis of Vps1 in a helix of the stalk domain identifies a mutant RR457-458EE that binds actin more weakly. In vivo analysis of Vps1 function demonstrates that the mutation disrupts endocytosis but not other functions of Vps1 such as vacuolar trafficking or peroxisome fission. The mutant Vps1 is stably expressed in cells and co-localizes with the endocytic reporters Abp1 and the amphiphysin Rvs167. Detailed analysis of individual endocytic patch behavior indicates that the mutation causes aberrant movements in later stages of endocytosis, consistent with a scission defect. Ultrastructural analysis of yeast cells using electron microscopy reveals a significant increase in invagination depth, further supporting a role for the Vps1-actin interaction during scission. In vitro analysis of the mutant protein demonstrates that—like wild-type Vps1—it is able to form oligomeric rings, but, critically, it has lost its ability to bundle actin filaments into higher-order structures. A model is proposed in which actin filaments bind Vps1 during invagination, and this interaction is important to transduce the force of actin polymerization to the membrane to drive successful scission. Yeast dynamin Vps1 forms oligomeric rings that bind and bundle F-actin in vitro Mutations in the Vps1 stalk region disrupt actin binding and bundling The Vps1 RR-EE actin binding mutant has defects in endocytic scission in vivo Palmer et al. reveal that the yeast dynamin Vps1 is able to form oligomeric rings that can directly interact with and bundle F-actin. Defects in the Vps1-actin interaction confer endocytic scission defects in vivo.
登录
查看更多内容
影响因子：
21.3
作者：
通讯作者：
--
影响因子：
64.5
作者：
Kaksonen, M;Toret, CP;Drubin, DG
通讯作者：
Drubin, DG
DOI：
10.1007/978-1-60761-783-9_15
发表时间：
2010-01-01
期刊：
IMMUNOELECTRON MICROSCOPY: METHODS AND PROTOCOLS
影响因子：
--
作者：
Fiserova, Jindriska;Goldberg, Martin W.
通讯作者：
Goldberg, Martin W.
影响因子：
21.3
作者：
Aghamohammadzadeh, Soheil;Ayscough, Kathryn R.
通讯作者：
Ayscough, Kathryn R.
影响因子：
4.5
作者：
Durieux, Anne-Cecile;Vassilopoulos, Stephane;Bitoun, Marc
通讯作者：
Bitoun, Marc