Nudel is crucial for the WAVE complex assembly in vivo by selectively promoting subcomplex stability and formation through direct interactions

Nudel is crucial for the WAVE complex assembly in vivo by selectively promoting subcomplex stability and formation through direct interactions
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Nudel 通过直接相互作用选择性地促进子复合物的稳定性和形成,对于体内 WAVE 复合物的组装至关重要

DOI:
10.1038/cr.2012.47
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发表时间:
2012-03
期刊:
影响因子:
44.1
通讯作者:
Zhu, Xueliang
Zhu, Xueliang
中科院分区:
生物学1区
文献类型:
--
作者:
Wu, Shuang;Ma, Li;Wu, Yibo;Zeng, Rong;Zhu, Xueliang

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由WAVE、Sra、Nap、Abi和HSPC 300组成的WAVE调节复合物(WRC)激活Arp 2/3复合物以响应Rac激活来控制分支肌动蛋白聚合。WRC如何在体内组装尚不清楚。在这里,我们表明,Nudel,一种对片状伪足形成至关重要的蛋白质,通过与Sra 1的动态结合,显着稳定了Sra 1-Nap 1-Abi 1复合物在细胞中的降解,而其与HSPC 300的物理相互作用保护游离HSPC 300免受蛋白酶体介导的降解,并刺激HSPC 300-WAVE 2复合物的形成。相比之下,Nudel与Sra 1-Nap 1-Abi 1-WAVE 2和Sra 1-Nap 1-Abi 1-HSPC 300复合物以及成熟WRC几乎没有相互作用。通过RNAi去除Nudel导致一般亚基降解并显著减弱成熟WRC的水平。它还取消了WRC依赖的肌动蛋白聚合在体外和Rac 1诱导的层状lipodial肌动蛋白网络的形成在细胞铺展。因此,Nudel通过拮抗某些WRC亚基和亚复合物的不稳定性,对于体内WRC组装的早期步骤是重要的。
The WAVE regulatory complex (WRC), consisting of WAVE, Sra, Nap, Abi, and HSPC300, activates the Arp2/3 complex to control branched actin polymerization in response to Rac activation. How the WRC is assembled in vivo is not clear. Here we show that Nudel, a protein critical for lamellipodia formation, dramatically stabilized the Sra1-Nap1-Abi1 complex against degradation in cells through a dynamic binding to Sra1, whereas its physical interaction with HSPC300 protected free HSPC300 from the proteasome-mediated degradation and stimulated the HSPC300-WAVE2 complex formation. By contrast, Nudel showed little or no interactions with the Sra1-Nap1-Abi1-WAVE2 and the Sra1-Nap1-Abi1-HSPC300 complexes as well as the mature WRC. Depletion of Nudel by RNAi led to general subunit degradation and markedly attenuated the levels of mature WRC. It also abolished the WRC-dependent actin polymerization in vitro and the Rac1-induced lamellipodial actin network formation during cell spreading. Therefore, Nudel is important for the early steps of the WRC assembly in vivo by antagonizing the instability of certain WRC subunits and subcomplexes.
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