Department of Neurology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA, USA.
Department of Pathology, University of Illinois College of Medicine, Chicago, IL, USA.
Cereb Cortex. 2019 May 1;29(5):1938-1952. doi: 10.1093/cercor/bhy073.
Although neural progenitor proliferation along the ventricular zone is regulated by β-catenin through Wnt signaling, the cytoskeletal mechanisms that regulate expression and localization of these proteins are not well understood. Our prior studies have shown that loss of the actin-binding Filamin A (FlnA) and actin-nucleating protein Formin 2 (Fmn2) impairs endocytosis of low-density-lipoprotein-receptor-related protein 6 (Lrp6), thereby disrupting β-catenin activation, resulting in decreased brain size. Here, we report that activated RhoA-GTPase disengages Fmn2 N- to C-terminal binding to promote Fmn2 activation and redistribution into lysosomal vesicles. Fmn2 colocalizes with β-catenin in lysosomes and promotes its degradation. Further, Fmn2 binds the E3 ligase Smurf2, enhances Smurf2-dependent ubiquitination, and degradation of Dishevelled-2 (Dvl2), thereby initiates β-catenin degradation. Finally, Fmn2 overexpression disrupts neuroepithelial integrity, neuronal migration, and proliferation-phenotypes in E13 mouse embryos, as seen with loss of Fmn2+FlnA function. Conversely, co-expression of Dvl2 with Fmn2 rescues the proliferation defect due to Fmn2 overexpression in mouse embryos. These findings suggest that there is a homeostatic feedback mechanism in the cytoskeletal-dependent regulation of neural proliferation within the cerebral cortex. Upstream, Fmn2 promotes proliferation by stabilizing the Lrp6 receptor, leading to β-catenin activation. Downstream, RhoA-activated Fmn2 promotes lysosomal degradation of Dvl2, leading to β-catenin degradation.
尽管神经祖细胞沿着脑室区的增殖受到β-连环蛋白通过 Wnt 信号的调节,但调节这些蛋白质表达和定位的细胞骨架机制尚不清楚。我们之前的研究表明,丧失肌动蛋白结合蛋白 Filamin A (FlnA) 和肌动蛋白成核蛋白 Formin 2 (Fmn2) 会损害低密度脂蛋白受体相关蛋白 6 (Lrp6) 的内吞作用,从而破坏β-连环蛋白的激活,导致大脑体积减小。在这里,我们报告说激活的 RhoA-GTPase 使 Fmn2 N-到 C-末端结合脱离,从而促进 Fmn2 的激活和再分布到溶酶体小泡中。Fmn2 与β-连环蛋白在溶酶体中共定位,并促进其降解。此外,Fmn2 与 E3 连接酶 Smurf2 结合,增强 Smurf2 依赖性泛素化和 Dishevelled-2 (Dvl2) 的降解,从而启动β-连环蛋白的降解。最后,Fmn2 的过表达破坏了 E13 天小鼠胚胎中的神经上皮完整性、神经元迁移和增殖表型,这与 Fmn2+FlnA 功能缺失的情况相同。相反,Dvl2 与 Fmn2 的共表达挽救了由于 Fmn2 过表达而导致的胚胎增殖缺陷。这些发现表明,在大脑皮层中,神经增殖的细胞骨架依赖性调节存在一种体内平衡反馈机制。在这一机制中,Fmn2 通过稳定 Lrp6 受体促进增殖,从而激活β-连环蛋白。在下游,RhoA 激活的 Fmn2 促进 Dvl2 的溶酶体降解,从而导致β-连环蛋白的降解。