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Updated: May 7, 2026
Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
Published on: May 3, 2018
CDK1 phosphorylation of YAP promotes mitotic defects and cell motility and is essential for neoplastic transformation
Shuping Yang1, Lin Zhang, Miao Liu
1Authors' Affiliations: Eppley Institute for Research in Cancer and Allied Diseases; Department of Pathology and Microbiology; and Mass Spectrometry and Proteomics Core Facility, University of Nebraska Medical Center, Omaha, Nebraska.
Abstract:
The Yes-associated protein, YAP, is a downstream effector of the Hippo pathway of cell-cycle control that plays important roles in tumorigenesis. Hippo-mediated phosphorylation YAP, mainly at S127, inactivates YAP function. In this study, we define a mechanism for positive regulation of YAP activity that is critical for its oncogenic function. Specifically, we found that YAP is phosphorylated in vitro and in vivo by the cell-cycle kinase CDK1 at T119, S289, and S367 during the G2-M phase of the cell cycle. We also found that ectopic expression of a phosphomimetic YAP mutant (YAP3D, harboring T119D/S289D/S367D) was sufficient to induce mitotic defects in immortalized epithelial cells, including centrosome amplification, multipolar spindles, and chromosome missegregation. Finally, we documented that mitotic phosphorylation of YAP was sufficient to promote cell migration and invasion in a manner essential for neoplastic cell transformation. In support of our findings, CDK1 inhibitors largely suppressed cell motility mediated by activated YAP-S127A but not the phosphomimetic mutant YAP3D. Collectively, our results reveal a previously unrecognized mechanism for controlling the activity of YAP that is crucial for its oncogenic function mediated by mitotic dysregulation.
Insights
The Yes-associated protein (YAP) is regulated by cell division kinase 1 (CDK1) during mitosis. This phosphorylation promotes cell migration and invasion, driving cancer development.
Area of Science:
- Cell Biology
- Molecular Oncology
- Cancer Research
Background:
- The Yes-associated protein (YAP) is a key regulator of cell proliferation and a downstream target of the Hippo pathway.
- YAP activity is typically inhibited by Hippo-mediated phosphorylation, primarily at serine 127 (S127).
- Dysregulation of YAP contributes significantly to tumorigenesis.
Purpose of the Study:
- To elucidate a novel mechanism for the positive regulation of YAP activity.
- To investigate the role of cell-cycle kinases in YAP activation during mitosis.
- To understand YAP's contribution to oncogenic functions through mitotic regulation.
Main Methods:
- In vitro and in vivo phosphorylation assays using cell-cycle kinase CDK1.
- Generation and analysis of a phosphomimetic YAP mutant (YAP3D).
- Assessment of mitotic defects, cell migration, and invasion in immortalized epithelial cells.
- Evaluation of CDK1 inhibitor effects on YAP-mediated cell motility.
Main Results:
- YAP is phosphorylated by CDK1 at specific sites (T119, S289, S367) during the G2-M phase.
- Ectopic expression of YAP3D induced mitotic abnormalities, including centrosome amplification and chromosome missegregation.
- Mitotic YAP phosphorylation enhanced cell migration and invasion, crucial for neoplastic transformation.
- CDK1 inhibitors reduced cell motility driven by YAP-S127A but not YAP3D.
Conclusions:
- CDK1-mediated phosphorylation of YAP during mitosis represents a novel mechanism for YAP activation.
- This mitotic regulation of YAP is critical for its oncogenic functions, particularly promoting cell motility and invasion.
- Targeting CDK1 may offer therapeutic strategies for cancers with YAP-driven oncogenesis.
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