Moreau V, Way M
Cell Biology Programme, European Molecular Biology Laboratory, Heidelberg, Germany.
FEBS Lett. 1998 May 15;427(3):353-6. doi: 10.1016/s0014-5793(98)00443-8.
In vitro actin based motility assays with bacterial pathogens have provided powerful systems to both understand and dissect actin dynamics as well as cell motility. Taking advantage of endogenous membrane vesicles in Xenopus extracts we have developed an in vitro assay to study membrane dependent actin polymerization. Our results demonstrate that membrane dependent actin polymerization, in contrast to Listeria stimulated actin filament assembly, is dependent on small GTPases of the Rho family. Using a combination of depletion and reconstitution experiments we have shown that Cdc42 but not Rac or Rho is required to stimulate actin polymerization from membranes. The in vitro system we have described here is amenable to identification of the downstream effectors of Cdc42 required for membrane dependent actin polymerization.
利用基于肌动蛋白的体外运动分析方法研究细菌病原体，为理解和剖析肌动蛋白动力学以及细胞运动提供了强大的系统。我们利用非洲爪蟾提取物中的内源性膜泡，开发了一种体外分析方法来研究膜依赖性肌动蛋白聚合。我们的结果表明，与李斯特菌刺激的肌动蛋白丝组装相反，膜依赖性肌动蛋白聚合依赖于Rho家族的小GTP酶。通过缺失和重组实验相结合，我们发现刺激膜上的肌动蛋白聚合需要Cdc42，而不是Rac或Rho。我们在此描述的体外系统适合鉴定膜依赖性肌动蛋白聚合所需的Cdc42的下游效应器。
