title: Phosphorylation of Tyr188 in the WW domain of YAP1 plays an essential role in YAP1-induced cellular transformation.
abstract: The Hippo signaling pathway regulates cellular proliferation and survival, thus exerting profound effects on normal cell fate and tumorigenesis. The pivotal effector of this pathway is YAP1, a transcriptional co-activator amplified in mouse and human cancers where it promotes epithelial-to-mesenchymal transition (EMT) and malignant transformation. The Hippo tumor suppressor pathway has been suggested to inhibit the YAP1 function through serine phosphorylation-induced cytoplasmic retention and degradation. Here we report that the tyrosine188 (Y188) site of YAP1 isoform with 2 WW domains (known as YAP1-2) plays an important role in YAP1-induced cellular transformation. IP-Mass Spectrometry analysis of YAP1 identified the phosphorylation of Y188 but not other tyrosine residues. In contrast to the aberrant 3D acinus formation observed in YAP1-WT transduced cells, overexpression of YAP1-Y188F (non-phosphorylated mimic) displayed normal 3D structures. In addition, knockdown of the endogenous YAP1 in MDA-MB231 breast cancer cells inhibited cell proliferation and migration, which were then successfully rescued by the exogenous YAP1-WT and YAP1-Y188E but not Y188F. Mechanistically, we also demonstrated that YAP1-Y188F had a higher affinity to the upstream negative regulator PTPN14 and was extensively localized in the cytoplasm. Since the Y188 is located in the conserved aromatic core of the WW domain of YAP1, our finding has a wide implication for WW domain signaling in general, where Y phosphorylation may act as a common positive regulator of the complex formation via WW domains. In summary, our results indicate that tyrosine 188 plays an important role in the YAP1-induced cellular transformation and its phosphorylation may intriguingly serve as a positive indicator of YAP1 activation.
authors.affiliation: a Department of Cancer Genetics , Roswell Park Cancer Institute , Buffalo , NY , USA.
authors.forename: Ying-Wei
authors.initials: YW
authors.lastname: Li
authors.affiliation: a Department of Cancer Genetics , Roswell Park Cancer Institute , Buffalo , NY , USA.
authors.forename: Jin
authors.initials: J
authors.lastname: Guo
authors.affiliation: a Department of Cancer Genetics , Roswell Park Cancer Institute , Buffalo , NY , USA.
authors.forename: He
authors.initials: H
authors.lastname: Shen
authors.affiliation: b Department of Pharmaceutical Sciences , New York Center of Excellence in Bioinformatics and Life Sciences, State University of New York , Buffalo , NY , USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.affiliation: a Department of Cancer Genetics , Roswell Park Cancer Institute , Buffalo , NY , USA.
authors.forename: Nuo
authors.initials: N
authors.lastname: Yang
authors.affiliation: a Department of Cancer Genetics , Roswell Park Cancer Institute , Buffalo , NY , USA.
authors.forename: Costa
authors.initials: C
authors.lastname: Frangou
authors.affiliation: a Department of Cancer Genetics , Roswell Park Cancer Institute , Buffalo , NY , USA.
authors.forename: Kayla E
authors.initials: KE
authors.lastname: Wilson
authors.affiliation: a Department of Cancer Genetics , Roswell Park Cancer Institute , Buffalo , NY , USA.|c Orthopaedic Oncology Institute, Tangdu Hospital, Fourth Military Medical University , Xi'an , Shaanxi , P. R. China.
authors.forename: Yinglong
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: a Department of Cancer Genetics , Roswell Park Cancer Institute , Buffalo , NY , USA.
authors.forename: Ashley L
authors.initials: AL
authors.lastname: Mussell
authors.affiliation: d Department of Physiology , National University of Singapore, The Yong Loo Li School of Medicine, Mechanobiology Institute, Institute of Molecular and Cell Biology (IMCB) A*STAR , Singapore , Republic of Singapore.
authors.forename: Marius
authors.initials: M
authors.lastname: Sudol
authors.affiliation: e Department of Biochemistry & Molecular Biology , Leonard Miller School of Medicine, University of Miami , Miami , FL , USA.
authors.forename: Amjad
authors.initials: A
authors.lastname: Farooq
authors.affiliation: b Department of Pharmaceutical Sciences , New York Center of Excellence in Bioinformatics and Life Sciences, State University of New York , Buffalo , NY , USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Qu
authors.affiliation: a Department of Cancer Genetics , Roswell Park Cancer Institute , Buffalo , NY , USA.
authors.forename: Jianmin
authors.initials: J
authors.lastname: Zhang
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry* / Q000378:metabolism*; D000595:Amino Acid Sequence; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D009024:Morphogenesis; D009857:Oncogenes; D010750:Phosphoproteins / Q000737:chemistry* / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism*; D000072417:Protein Domains; D011499:Protein Processing, Post-Translational; D014157:Transcription Factors; D000091102:YAP-Signaling Proteins
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D010750:Phosphoproteins; D014157:Transcription Factors; D000091102:YAP-Signaling Proteins; C088374:YAP1 protein, human; D019000:Phosphotyrosine
keywords: Hippo pathway; YAP1; cellular transformation; tyrosine phosphorylation