Role of Angiomotin-like 2 mono-ubiquitination on YAP inhibition
Role of Angiomotin-like 2 mono-ubiquitination on YAP inhibition
复制标题
DOI：
10.15252/embr.201540809
复制
发表时间：
2016-01-01
期刊：
影响因子：
7.7
通讯作者：
Lim, Dae-Sik
中科院分区：
文献类型：
--
作者：
Kim, Miju;Kim, Minchul;Lim, Dae-Sik
LATS1/2（大肿瘤抑制因子）激酶和Angiomotin家族蛋白是YAP （yes-associated protein）癌蛋白的有效抑制剂，但其潜在的分子机制尚不完全清楚。在这里，我们首次报道了USP9X是血管运动素样2 （AMOTL2）的去泛素酶，并且AMOTL2的单泛素化是抑制YAP所必需的。USP9X敲低增加了lats介导的YAP磷酸化，减少了YAP的转录输出。相反，在密集培养的细胞中，USP9X的过表达会重新激活YAP。遗传和生化方法都确定AMOTL2是USP9X的靶标。AMOTL2被发现在K347和K408位点泛素化，这两个位点都位于蛋白质的螺旋结构域。AMOTL2 K347/408R突变体不能被泛素化，其抑制YAP的能力受损。此外，泛素化的AMOTL2可以结合到LATS激酶的UBA结构域，而该结构域是LATS功能所必需的。我们的研究结果为核心Hippo通路组分的激活机制提供了新的见解。
LATS1/2 (large tumor suppressor) kinases and the Angiomotin family proteins are potent inhibitors of the YAP (yes-associated protein) oncoprotein, but the underlying molecular mechanism is not fully understood. Here, we report for the first time that USP9X is a deubiquitinase of Angiomotin-like 2 (AMOTL2) and that AMOTL2 mono-ubiquitination is required for YAP inhibition. USP9X knockdown increased the LATS-mediated phosphorylation of YAP and decreased the transcriptional output of YAP. Conversely, overexpression of USP9X reactivated YAP in densely cultured cells. Both genetic and biochemical approaches identified AMOTL2 as a target of USP9X. AMOTL2 was found to be ubiquitinated at K347 and K408, which both reside in the protein's coiled-coil domain. The AMOTL2 K347/408R mutant, which cannot be ubiquitinated, was impaired in its ability to inhibit YAP. Furthermore, ubiquitinated AMOTL2 can bind to the UBA domain of LATS kinase, and this domain is required for the function of LATS. Our results provide novel insights into the activation mechanisms of core Hippo pathway components.