title: Role of Angiomotin-like 2 mono-ubiquitination on YAP inhibition.
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.
authors.affiliation: National Creative Research Center for Cell Division and Differentiation, Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Korea.
authors.forename: Miju
authors.initials: M
authors.lastname: Kim
authors.affiliation: National Creative Research Center for Cell Division and Differentiation, Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Korea.
authors.forename: Minchul
authors.initials: M
authors.lastname: Kim
authors.affiliation: Center for Theragnosis, Biomedical Research Institute Korea Institute of Science and Technology (KIST), Seoul, Korea.
authors.forename: Seong-Jun
authors.initials: SJ
authors.lastname: Park
authors.affiliation: Center for Theragnosis, Biomedical Research Institute Korea Institute of Science and Technology (KIST), Seoul, Korea Department of Biological Chemistry, University of Science and Technology, Daejeon, Korea.
authors.forename: Cheolju
authors.initials: C
authors.lastname: Lee
authors.affiliation: National Creative Research Center for Cell Division and Differentiation, Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Korea daesiklim@kaist.ac.kr.
authors.forename: Dae-Sik
authors.initials: DS
authors.lastname: Lim
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics* / Q000378:metabolism; D000091082:Angiomotins; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008840:Microfilament Proteins; D015513:Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics* / Q000378:metabolism; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins / Q000378:metabolism; D043222:Ubiquitin Thiolesterase / Q000235:genetics; D054875:Ubiquitination; D000091102:YAP-Signaling Proteins
chemical_list: C423985:AMOT protein, human; D048868:Adaptor Proteins, Signal Transducing; D000091082:Angiomotins; D002352:Carrier Proteins; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D008840:Microfilament Proteins; D015513:Oncogene Proteins; D010750:Phosphoproteins; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C506234:USP9X protein, human; D000091102:YAP-Signaling Proteins; C088374:YAP1 protein, human; C092745:LATS1 protein, human; D017346:Protein Serine-Threonine Kinases; D043222:Ubiquitin Thiolesterase
keywords: AMOTL2; Hippo pathway; LATS‐YAP; USP9X; mono‐ubiquitination