Selective targeting of activating and inhibitory Smads by distinct WWP2 ubiquitin ligase isoforms differentially modulates TGFβ signalling and EMT.
Selective targeting of activating and inhibitory Smads by distinct WWP2 ubiquitin ligase isoforms differentially modulates TGFβ signalling and EMT.
复制标题
通过不同的 WWP2 泛素连接酶异构体选择性靶向激活和抑制性 Smad，差异调节 TGFβ 信号传导和 EMT。
DOI：
10.1038/onc.2010.617
复制
发表时间：
2011-05-26
期刊：
影响因子：
8
通讯作者：
Chantry, A.
中科院分区：
文献类型：
--
作者：
Soond, S. M.;Chantry, A.
泛素依赖性机制已成为控制Smads和TGFβ依赖性生物输出（如上皮-间充质转化（EMT））的细胞水平的重要调控元件。在这里，我们确定了一种HECT E3泛素连接酶，称为WWP 2（全长WWP 2-FL），连同两个WWP 2亚型（N-末端，WWP 2-N; C-末端WWP 2-C），作为新的Smad结合伴侣。我们发现WWP 2-FL在TGFβ通路中只与Smad 2、Smad 3和Smad 7相互作用。有趣的是，WWP 2-N亚型与Smad 2和Smad 3相互作用，而WWP 2-C仅与Smad 7相互作用。此外，基于蛋白质周转和泛素化研究，WWP 2-FL和WWP 2-C具有对Smad 7的偏好。出乎意料的是，我们还发现缺乏HECT泛素连接酶结构域的WWP 2-N也可以以TGFβ调节的方式与WWP 2-FL相互作用，并激活内源性WWP 2泛素连接酶活性，导致未受刺激的Smad 2和Smad 3降解。与我们的蛋白质相互作用数据一致，过表达和敲低方法揭示WWP 2同种型差异性地调节TGFβ依赖性转录和EMT。最后，我们发现选择性破坏WWP 2与抑制性Smad 7的相互作用可以稳定Smad 7蛋白水平并防止TGFβ诱导的EMT。总的来说，我们的数据表明，WWP 2-N可以刺激WWP 2-FL，导致针对未受刺激的Smad 2和Smad 3的活性增加，并且Smad 7是延长的TGFβ刺激后WWP 2-FL和WWP 2-C的优选底物。值得注意的是，这是第一份关于不同HECT E3遍在蛋白连接酶亚型相互依赖的生物学作用的报告，并强调了一种全新的调节范式，可以选择性地限制抑制性和激活性Smads的水平。
Ubiquitin-dependent mechanisms have emerged as essential regulatory elements controlling cellular levels of Smads and TGFβ-dependent biological outputs such as epithelial-mesenchymal transition (EMT). Here, we identify a HECT E3 ubiquitin ligase known as WWP2 (Full-length WWP2-FL), together with two WWP2 isoforms (N-terminal, WWP2-N; C-terminal WWP2-C), as novel Smad binding partners. We show that WWP2-FL interacts exclusively with Smad2, Smad3 and Smad7 in the TGFβ pathway. Interestingly, the WWP2-N isoform interacts with Smad2 and Smad3, whereas WWP2-C interacts only with Smad7. In addition, WWP2-FL and WWP2-C have a preference for Smad7 based on protein turnover and ubiquitination studies. Unexpectedly, we also find that WWP2-N, which lacks the HECT ubiquitin ligase domain, can also interact with WWP2-FL in a TGFβ-regulated manner and activate endogenous WWP2 ubiquitin ligase activity causing degradation of unstimulated Smad2 and Smad3. Consistent with our protein interaction data, overexpression and knockdown approaches reveal that WWP2 isoforms differentially modulate TGFβ-dependent transcription and EMT. Finally, we show that selective disruption of WWP2 interactions with inhibitory Smad7 can stabilise Smad7 protein levels and prevent TGFβ-induced EMT. Collectively, our data suggest that WWP2-N can stimulate WWP2-FL leading to increased activity against unstimulated Smad2 and Smad3, and that Smad7 is a preferred substrate for WWP2-FL and WWP2-C following prolonged TGFβ stimulation. Significantly, this is the first report of an inter-dependent biological role for distinct HECT E3 ubiquitin ligase isoforms, and highlights an entirely novel regulatory paradigm that selectively limits the level of inhibitory and activating Smads.
登录
查看更多内容
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
3.7
作者：
Li W;Bengtson MH;Ulbrich A;Matsuda A;Reddy VA;Orth A;Chanda SK;Batalov S;Joazeiro CA
通讯作者：
Joazeiro CA
DOI：
10.1016/0922-3371(90)90079-c
发表时间：
1990-05-01
期刊：
CELL DIFFERENTIATION AND DEVELOPMENT
影响因子：
--
作者：
ATSUMI, T;MIWA, Y;IKAWA, Y
通讯作者：
IKAWA, Y
影响因子：
5.3
作者：
Li, Hui;Zhang, Zhihong;Jin, Ying
通讯作者：
Jin, Ying
DOI：
10.1006/bbrc.2001.6206
发表时间：
2002-01-11
影响因子：
3.1
作者：
Flasza, M;Gorman, P;Baron, M
通讯作者：
Baron, M