WASH and the Arp2/3 complex regulate endosome shape and trafficking.
WASH and the Arp2/3 complex regulate endosome shape and trafficking.
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DOI：
10.1002/cm.20437
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发表时间：
2010-03
期刊：
影响因子：
2.9
通讯作者：
Welch, Matthew D.
中科院分区：
文献类型：
--
作者：
Duleh, Steve N.;Welch, Matthew D.
Arp 2/3复合物的激活剂，称为成核促进因子（NPFs），是细胞中肌动蛋白组装的适当空间和时间控制所必需的。哺乳动物细胞表达几种NPF，每种NPF在不同的细胞过程中起作用，包括吞噬和内吞作用中的WASP和N-WASP，细胞迁移中的WAVE和JMY，以及ER至高尔基体运输中的WHAMM。虽然最近发现了另一种称为WASH的NPF，但该蛋白的细胞定位和功能尚不清楚。在这里，我们证明了人类WASH单独在体外和细胞中有效地激活Arp 2/3复合物，表明该蛋白质不像N-WASP那样被自抑制，但可能受到相互作用蛋白质的调节。在细胞中，WASH与Rab 5阳性早期内体和Rab 11阳性回收内体相关，这些内体富含肌动蛋白丝。通过RNAi沉默WASH或Arp 2/3复合物的表达，或通过药物处理破坏肌动蛋白功能，引起内体的扩大和延长。有趣的是，WASH沉默以及肌动蛋白破坏延迟了EGF转运到LAMP 1阳性晚期内体。这些观察结果表明，肌动蛋白聚合WASH影响的形状和成熟的内体，并突出了以前未被认识的作用WASH和Arp 2/3复合物的内吞运输的降解步骤。
Activators of the Arp2/3 complex, termed nucleation-promoting factors (NPFs), are required for the proper spatial and temporal control of actin assembly in cells. Mammalian cells express several NPFs, each of which functions in a distinct cellular process, including WASP and N-WASP in phagocytosis and endocytosis, WAVE and JMY in cell migration, and WHAMM in ER-to-Golgi transport. Although another NPF called WASH was recently identified, the cellular localization and function of this protein were unclear. Here we demonstrated that human WASH alone potently activated the Arp2/3 complex in vitro and in cells, suggesting that the protein is not autoinhibited like N-WASP, but is likely regulated by interacting proteins. In cells, WASH was associated with Rab5-positive early endosomes and Rab11-positive recycling endosomes that were enriched for actin filaments. Silencing of WASH or Arp2/3 complex expression by RNAi, or disruption of actin function by drug treatments, caused enlargement and elongation of endosomes. Intriguingly, WASH silencing, as well as actin disruption, delayed EGF transport to LAMP1-positive late endosomes. These observations indicate that actin polymerization by WASH influences the shape and maturation of endosomes, and highlight a previously unrecognized role for WASH and the Arp2/3 complex in the degradative steps of endocytic trafficking.
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影响因子：
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作者：
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