Department of Biology, York University, Toronto, Ontario M3J 1P3, Canada.
J Biol Chem. 2010 Jul 23;285(30):23040-6. doi: 10.1074/jbc.M109.096164. Epub 2010 May 25.
The Rho GTPase members and their effector proteins, such as the Wiskott-Aldrich syndrome protein (WASP), play critical roles in regulating actin dynamics that affect cell motility, endocytosis, cell division, and transport. It is well established that Caenorhabditis elegans wsp-1 plays an essential role in embryonic development. We were interested in the role of the C. elegans protein WSP-1 in the adult nematode. In this report, we show that a deletion mutant of wsp-1 exhibits a strong sensitivity to the neuromuscular inhibitor aldicarb. Transgenic rescue experiments demonstrated that neuronal expression of WSP-1 rescued this phenotype and that it required a functional WSP-1 Cdc42/Rac interactive binding domain. WSP-1-GFP fusion protein was found localized presynaptically, immediately adjacent to the synaptic protein RAB-3. Strong genetic interactions with wsp-1 and other genes involved in different stages of synaptic transmission were observed as the wsp-1(gm324) mutation suppresses the aldicarb resistance seen in unc-13(e51), unc-11(e47), and snt-1 (md290) mutants. These results provide genetic and pharmacological evidence that WSP-1 plays an essential role to stabilize the actin cytoskeleton at the neuronal active zone of the neuromuscular junction to restrain synaptic vesicle release.
Rho GTPase 成员及其效应蛋白,如 Wiskott-Aldrich 综合征蛋白 (WASP),在调节肌动蛋白动力学方面发挥着关键作用,肌动蛋白动力学影响细胞运动、内吞作用、细胞分裂和运输。已经证实,秀丽隐杆线虫 wsp-1 在胚胎发育中起着重要作用。我们对成年线虫中 C. elegans 蛋白 WSP-1 的作用感兴趣。在本报告中,我们表明 wsp-1 的缺失突变体对神经肌肉抑制剂 aldicarb 表现出强烈的敏感性。转基因拯救实验表明,神经元表达的 WSP-1 挽救了这种表型,并且它需要一个功能性的 WSP-1 Cdc42/Rac 相互作用结合域。WSP-1-GFP 融合蛋白被发现定位于突触前,紧邻突触蛋白 RAB-3。观察到与 wsp-1 和其他参与突触传递不同阶段的基因的强烈遗传相互作用,因为 wsp-1(gm324)突变抑制了 unc-13(e51)、unc-11(e47)和 snt-1 (md290)突变体中观察到的 aldicarb 抗性。这些结果提供了遗传和药理学证据,表明 WSP-1 在稳定神经元活性区的肌动蛋白细胞骨架方面发挥着重要作用,以限制突触囊泡释放。