The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan 5290002, Israel.
Sci Rep. 2017 Mar 23;7:44863. doi: 10.1038/srep44863.
WASp family Verprolin-homologous protein-2 (WAVE2), a member of the Wiskott-Aldrich syndrome protein (WASp) family of actin nucleation promoting factors, is a central regulator of actin cytoskeleton polymerization and dynamics. Multiple signaling pathways operate via WAVE2 to promote the actin-nucleating activity of the actin-related protein 2/3 (Arp2/3) complex. WAVE2 exists as a part of a pentameric protein complex known as the WAVE regulatory complex (WRC), which is unstable in the absence of its individual proteins. While the involvement of WAVE2 in actin polymerization has been well documented, its negative regulation mechanism is poorly characterized to date. Here, we demonstrate that WAVE2 undergoes ubiquitylation in a T-cell activation dependent manner, followed by proteasomal degradation. The WAVE2 ubiquitylation site was mapped to lysine 45, located at the N-terminus where WAVE2 binds to the WRC. Using Förster resonance energy transfer (FRET), we reveal that the autoinhibitory conformation of the WRC maintains the stability of WAVE2 in resting cells; the release of autoinhibition following T-cell activation facilitates the exposure of WAVE2 to ubiquitylation, leading to its degradation. The dynamic conformational structures of WAVE2 during cellular activation dictate its degradation.
WASp 家族中的 Verprolin 同源蛋白-2(WAVE2)是肌动蛋白成核促进因子 Wiskott-Aldrich 综合征蛋白(WASp)家族的成员,是肌动蛋白细胞骨架聚合和动力学的核心调节剂。多种信号通路通过 WAVE2 作用,促进肌动蛋白相关蛋白 2/3(Arp2/3)复合物的肌动蛋白成核活性。WAVE2 作为五聚体蛋白复合物 WAVE 调节复合物(WRC)的一部分存在,其在缺乏其单个蛋白时不稳定。尽管 WAVE2 参与肌动蛋白聚合已得到充分证实,但迄今为止,其负调控机制仍知之甚少。在这里,我们证明 WAVE2 以依赖 T 细胞激活的方式发生泛素化,随后被蛋白酶体降解。WAVE2 的泛素化位点被映射到赖氨酸 45,位于 N 端,WAVE2 在此处与 WRC 结合。使用Förster 共振能量转移(FRET),我们揭示了 WRC 的自抑制构象在静止细胞中保持 WAVE2 的稳定性;T 细胞激活后自抑制的释放促进了 WAVE2 暴露于泛素化,导致其降解。细胞激活过程中 WAVE2 的动态构象结构决定了其降解。