Role of lipids and actin in the formation of clathrin-coated pits
Role of lipids and actin in the formation of clathrin-coated pits
复制标题
DOI：
10.1016/j.yexcr.2006.09.025
复制
发表时间：
2006-12-10
影响因子：
3.7
通讯作者：
Ehrlich, Marcelo
中科院分区：
文献类型：
--
作者：
Boucrot, Emmanuel;Saffarian, Saveez;Ehrlich, Marcelo
网格蛋白包被小窝的组装及其成熟为包被囊泡需要特定脂质与几种结构和调节蛋白之间的协同相互作用。在伯醇存在的情况下，磷脂酶D产生磷脂酰醇而非磷脂酸（PA），减少了对磷脂酰肌醇5 - 激酶（PI5K）的刺激，从而降低了磷脂酰肌醇 - 4,5 - 二磷酸（PIP2）的形成。通过活细胞成像，我们已经表明，用1 - 丁醇或其他小分子伯醇对细胞进行急性处理会诱导质膜上包被小窝的快速解体，并阻止新的包被小窝出现。添加外源性PIP2可逆转这种效应。去除1 - 丁醇后，包被小窝和囊泡会同步重新出现；我们利用这种同步性来评估肌动蛋白在包被囊泡组装中的作用。用拉特伦菌素A或细胞松弛素D长时间抑制肌动蛋白聚合，会使包被小窝形成的频率降低约50%，但不影响其成熟为包被囊泡。与对照细胞一样，在肌动蛋白解聚剂持续存在的情况下去除1 - 丁醇会导致新的小窝同步出现，且这些小窝正常成熟。因此，肌动蛋白细胞骨架的重塑对于网格蛋白包被囊泡的组装不是必需的，但可能间接影响网格蛋白包被小窝的成核。（c）2006爱思唯尔公司。保留所有权利。
Assembly of clathrin-coated pits and their maturation into coated vesicles requires coordinated interactions between specific lipids and several structural and regulatory proteins. In the presence of primary alcohols, phospholipase D generates phosphatidylalcohols instead of PA, reducing stimulation of phosphatidyl inositol 5-kinase (PI5K) and hence decreasing formation of phosphoinositide-4,5-biphosphate (PIP2). Using live-cell imaging, we have shown that acute treatment of cells with 1-butanol or other small primary alcohols induces rapid disassembly of coated pits at the plasma membrane and blocks appearance of new ones. Addition of exogenous PIP2 reverses this effect. Coated pits and vesicles reappear synchronously upon removal of 1-butanol; we have used this synchrony to assess the role of actin in coated vesicle assembly. Prolonged inhibition of actin polymerization by latrunculin A or cytochalasin D reduced by similar to 50% the frequency of coated pit formation without affecting maturation into coated vesicles. As in control cells, removal of 1-butanol in the continued presence of an actin depolymerizer led to synchronous appearance of new pits, which matured normally. Thus, remodeling of the actin cytoskeleton is not essential for clathrin-coated vesicle assembly but may indirectly affect the nucleation of clathrin-coated pits. (c) 2006 Elsevier Inc. All rights reserved.