Miki H, Suetsugu S, Takenawa T
Department of Biochemistry, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108, Japan.
EMBO J. 1998 Dec 1;17(23):6932-41. doi: 10.1093/emboj/17.23.6932.
Rac is a Rho-family small GTPase that induces the formation of membrane ruffles. However, it is poorly understood how Rac-induced reorganization of the actin cytoskeleton, which is essential for ruffle formation, is regulated. Here we identify a novel Wiskott-Aldrich syndrome protein (WASP)-family protein, WASP family Verprolin-homologous protein (WAVE), as a regulator of actin reorganization downstream of Rac. Ectopically expressed WAVE induces the formation of actin filament clusters that overlap with the expressed WAVE itself. In this actin clustering, profilin, a monomeric actin-binding protein that has been suggested to be involved in actin polymerization, was shown to be essential. The expression of a dominant-active Rac mutant induces the translocation of endogenous WAVE from the cytosol to membrane ruffling areas. Furthermore, the co-expression of a deltaVPH WAVE mutant that cannot induce actin reorganization specifically suppresses the ruffle formation induced by Rac, but has no effect on Cdc42-induced actin-microspike formation, a phenomenon that is also known to be dependent on rapid actin reorganization. The deltaVPH WAVE also suppresses membrane-ruffling formation induced by platelet-derived growth factor in Swiss 3T3 cells. Taken together, we conclude that WAVE plays a critical role downstream of Rac in regulating the actin cytoskeleton required for membrane ruffling.
Rac是一种Rho家族小GTP酶,可诱导膜皱褶的形成。然而,对于Rac诱导的肌动蛋白细胞骨架重排(这对皱褶形成至关重要)是如何被调控的,人们了解甚少。在此,我们鉴定出一种新型的威斯科特-奥尔德里奇综合征蛋白(WASP)家族蛋白,即WASP家族维普洛林同源蛋白(WAVE),它是Rac下游肌动蛋白重排的调节因子。异位表达的WAVE可诱导肌动蛋白丝簇的形成,这些丝簇与表达的WAVE本身重叠。在这种肌动蛋白聚集过程中,原肌球蛋白(一种已被认为参与肌动蛋白聚合的单体肌动蛋白结合蛋白)被证明是必不可少的。显性活性Rac突变体的表达可诱导内源性WAVE从胞质溶胶转运至膜皱褶区域。此外,不能诱导肌动蛋白重排的deltaVPH WAVE突变体的共表达可特异性抑制Rac诱导的皱褶形成,但对Cdc42诱导的肌动蛋白微刺形成没有影响,Cdc42诱导的肌动蛋白微刺形成也是一种依赖于快速肌动蛋白重排的现象。deltaVPH WAVE还可抑制瑞士3T3细胞中血小板衍生生长因子诱导的膜皱褶形成。综上所述,我们得出结论,WAVE在Rac下游发挥关键作用,调控膜皱褶所需的肌动蛋白细胞骨架。