Role of Angiomotin‐like 2 mono‐ubiquitination on YAP inhibition. (23rd November 2015)

• Record Type: Journal Article
• Authors: Kim, Miju; Kim, Minchul; Park, Seong‐Jun; Lee, Cheolju; Lim, Dae‐Sik
• Is Part Of: EMBO reports. Volume 17:Number 1(2016:Jan.)
• Page Start: 64
• Page End: 78
• Publication Date: 2015-11-23
• Subjects: AMOTL2 -- Hippo pathway -- LATS‐YAP -- mono‐ubiquitination -- USP9X; Molecular biology -- Periodicals; Molecular Biology -- Periodicals; Molecular biology; Periodicals; 572.8
• DOI: 10.15252/embr.201540809
• Languages: English
• ISSNs: 1469-221X

Abstract

Abstract: 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, over‐expression 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. Synopsis: This study reports a new regulatory mechanism of the Hippo signaling governed by mono‐ubiquitination of AMOTL2. AMOTL2 mono‐ubiquitination is required to bind and activate LATS kinase and to inhibit YAP activity and is regulated by the USP9X deubiquitinase. USP9X is a deubiquitinating enzyme activating YAP. USP9X deubiquitinates AMOTL2 on its coiled‐coil domain. Mono‐ubiquitination of AMOTL2 is required for its tumor suppressor function. Mono‐ubiquitinated AMOTL2 activates LATS kinase through binding to the UBA domain of LATS. Abstract : This study reports a new regulatory mechanism of the Hippo signaling governed by mono‐ubiquitination of AMOTL2. AMOTL2 mono‐ubiquitination is required to bind and activate LATS kinase and to inhibit YAP activity and is regulated by the USP9X deubiquitinase.