Machesky L M, Insall R H
MRC-LMCB, Department of Molecular Medicine, University College London, Gower Street, London WC1E 6BT, UK.
Curr Biol. 1998;8(25):1347-56. doi: 10.1016/s0960-9822(98)00015-3.
The actin-related proteins Arp2 and Arp3 are part of a seven-protein complex which is localized in the lamellipodia of a variety of cell types, and in actin-rich spots of unknown function. The Arp2/3 complex enhances actin nucleation and causes branching and crosslinking of actin filaments in vitro; in vivo it is thought to drive the formation of lamellipodia and to be a control center for actin-based motility. The Wiskott-Aldrich syndrome protein, WASP, is an adaptor protein implicated in the transmission of signals from tyrosine kinase receptors and small GTPases to the actin cytoskeleton. Scar1 is a member of a new family of proteins related to WASP, and it may also have a role in regulating the actin cytoskeleton. Scar1 is the human homologue of Dictyostelium Scar1, which is thought to connect G-protein-coupled receptors to the actin cytoskeleton. The mammalian Scar family contains at least four members. We have examined the relationships between WASP, Scar1, and the Arp2/3 complex.
We have identified WASP and its relative Scar1 as proteins that interact with the Arp2/3 complex. We have used deletion analysis to show that both WASP and Scar1 interact with the p21 subunit of the Arp2/3 complex through their carboxyl termini. Overexpression of carboxy-terminal fragments of Scar1 or WASP in cells caused a disruption in the localization of the Arp2/3 complex and, concomitantly, induced a complete loss of lamellipodia and actin spots. The induction of lamellipodia by platelet-derived growth factor was also suppressed by overexpression of the fragment of Scar1 that binds to the Arp2/3 complex.
We have identified a conserved sequence domain in proteins of the WASP family that binds to the Arp2/3 complex. Overexpression of this domain in cells disrupts the localization of the Arp2/3 complex and inhibits lamellipodia formation. Our data suggest that WASP-related proteins may regulate the actin cytoskeleton through the Arp2/3 complex.
肌动蛋白相关蛋白Arp2和Arp3是一个七蛋白复合物的组成部分,该复合物定位于多种细胞类型的板状伪足以及功能未知的富含肌动蛋白的位点。Arp2/3复合物在体外可增强肌动蛋白成核作用,并导致肌动蛋白丝分支和交联;在体内,它被认为可驱动板状伪足的形成,并且是基于肌动蛋白的运动的控制中心。威斯科特-奥尔德里奇综合征蛋白(WASP)是一种衔接蛋白,参与从酪氨酸激酶受体和小GTP酶向肌动蛋白细胞骨架的信号传递。Scar1是与WASP相关的一个新蛋白家族的成员,它可能也在调节肌动蛋白细胞骨架中发挥作用。Scar1是盘基网柄菌Scar1的人类同源物,后者被认为可将G蛋白偶联受体与肌动蛋白细胞骨架相连。哺乳动物Scar家族至少包含四个成员。我们已经研究了WASP、Scar1与Arp2/3复合物之间的关系。
我们已鉴定出WASP及其相关蛋白Scar1是与Arp2/3复合物相互作用的蛋白。我们通过缺失分析表明,WASP和Scar1均通过其羧基末端与Arp2/3复合物的p21亚基相互作用。在细胞中过表达Scar1或WASP的羧基末端片段会导致Arp2/3复合物的定位紊乱,并随之导致板状伪足和肌动蛋白位点完全丧失。与Arp2/3复合物结合的Scar1片段的过表达也会抑制血小板衍生生长因子诱导的板状伪足形成。
我们已在WASP家族蛋白中鉴定出一个与Arp2/3复合物结合的保守序列结构域。该结构域在细胞中的过表达会破坏Arp2/3复合物的定位并抑制板状伪足的形成。我们的数据表明,WASP相关蛋白可能通过Arp2/3复合物调节肌动蛋白细胞骨架。