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NEK1 Phosphorylation of YAP Promotes Its Stabilization and Transcriptional Output

Md Imtiaz Khalil1, Ishita Ghosh1, Vibha Singh1

  • 1Department of Biochemistry and Molecular Biology, LSU Health Sciences Center, Shreveport, LA 71130, USA.

Cancers
|December 10, 2020
PubMed

Insights

A new TLK1>NEK1>YAP1 axis drives prostate cancer progression. This pathway promotes androgen-independent growth, a key step in lethal metastatic castration-resistant prostate cancer (mCRPC).

Area of Science:

  • Oncology
  • Molecular Biology
  • Cell Biology

Background:

  • Prostate cancer (PCa) deaths often stem from treatment failure, leading to metastatic castration-resistant PCa (mCRPC).
  • Mechanisms driving resistance to androgen deprivation therapy (ADT) remain incompletely understood.
  • The roles of tousled-like kinase 1 (TLK1) and never in mitosis gene A (NIMA)-related kinase 1 (NEK1) in DNA damage response (DDR) and cell cycle arrest in PCa cells were investigated.

Purpose of the Study:

  • To elucidate the role of TLK1, NEK1, and YAP1 in the transition to androgen-independent (AI) growth in prostate cancer.
  • To identify the molecular interactions and signaling axis governing AI growth adaptation.
  • To explore the clinical relevance of the NEK1-YAP1 interaction in various cancers.

Main Methods:

  • Overexpression of wild-type NEK1 (wt-NEK1) in LNCaP cells to observe effects on androgen independence.
  • Co-immunoprecipitation assays to detect NEK1-YAP1 interaction.
  • In vitro kinase assays to determine NEK1's phosphorylation of YAP1.
  • NEK1 knockout (KO) in NT1 PCa cells to assess YAP1 levels and target gene expression.
  • Analysis of NEK1 and YAP1 protein expression in clinical cancer samples.

Main Results:

  • Overexpression of wt-NEK1 induced rapid androgen-independent (AI) growth, similar to YAP1 overexpression.
  • wt-NEK1 overexpression led to YAP1 accumulation, suggesting a TLK1>NEK1>YAP1 signaling axis.
  • NEK1 directly phosphorylated YAP1 on six residues in vitro, likely stabilizing the protein.
  • NEK1 KO reduced YAP1 levels and YAP-regulated gene expression in PCa cells.
  • NEK1 and YAP1 expression were concurrently elevated and correlated in prostate cancer, head and neck squamous cell carcinoma, and glioblastoma.

Conclusions:

  • The TLK1>NEK1>YAP1 axis is identified as a critical pathway promoting AI growth and cancer progression.
  • NEK1-mediated YAP1 phosphorylation and stabilization are key events in the androgen-sensitive to androgen-independent transition in PCa.
  • The NEK1-YAP1 co-regulation has significant implications for understanding and potentially targeting mCRPC and other cancers.
Keywords:
MS-determined phosphopeptidesNIMA-related kinase 1 (NEK1)thioridazine (THD)tousled-like kinase (TLK)yes-associated protein 1 (YAP1)

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