Role of Angiomotin-like 2 mono-ubiquitination on YAP inhibition

Miju Kim; Minchul Kim; Seong-Jun Park; Cheolju Lee; Dae-Sik Lim

doi:10.15252/embr.201540809

Role of Angiomotin-like 2 mono-ubiquitination on YAP inhibition

EMBO Rep. 2016 Jan;17(1):64-78. doi: 10.15252/embr.201540809. Epub 2015 Nov 23.

Authors

Miju Kim¹, Minchul Kim¹, Seong-Jun Park², Cheolju Lee³, Dae-Sik Lim⁴

Affiliations

¹ National Creative Research Center for Cell Division and Differentiation, Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Korea.
² Center for Theragnosis, Biomedical Research Institute Korea Institute of Science and Technology (KIST), Seoul, Korea.
³ 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.
⁴ 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.

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.

Keywords: AMOTL2; Hippo pathway; LATS‐YAP; USP9X; mono‐ubiquitination.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adaptor Proteins, Signal Transducing / genetics*
Adaptor Proteins, Signal Transducing / metabolism
Angiomotins
Carrier Proteins / genetics
Carrier Proteins / metabolism
HEK293 Cells
Humans
Intercellular Signaling Peptides and Proteins / genetics
Intercellular Signaling Peptides and Proteins / metabolism*
Membrane Proteins / genetics
Membrane Proteins / metabolism*
Microfilament Proteins
Oncogene Proteins / genetics
Oncogene Proteins / metabolism
Phosphoproteins / genetics*
Phosphoproteins / metabolism
Phosphorylation
Protein Serine-Threonine Kinases / metabolism
Signal Transduction / genetics
Transcription Factors
Tumor Suppressor Proteins / metabolism
Ubiquitin Thiolesterase / genetics
Ubiquitination
YAP-Signaling Proteins

Substances

AMOT protein, human
Adaptor Proteins, Signal Transducing
Angiomotins
Carrier Proteins
Intercellular Signaling Peptides and Proteins
Membrane Proteins
Microfilament Proteins
Oncogene Proteins
Phosphoproteins
Transcription Factors
Tumor Suppressor Proteins
USP9X protein, human
YAP-Signaling Proteins
YAP1 protein, human
LATS1 protein, human
Protein Serine-Threonine Kinases
Ubiquitin Thiolesterase