A cell-based screening for TAZ activators identifies ethacridine, a widely used antiseptic and abortifacient, as a compound that promotes dephosphorylation of TAZ and inhibits adipogenesis in C3H10T1/2 cells

Shodai Kawano; Junichi Maruyama; Shunta Nagashima; Kazutoshi Inami; Wenzhe Qiu; Hiroaki Iwasa; Kentaro Nakagawa; Mari Ishigami-Yuasa; Hiroyuki Kagechika; Hiroshi Nishina; Yutaka Hata

doi:10.1093/jb/mvv051

A cell-based screening for TAZ activators identifies ethacridine, a widely used antiseptic and abortifacient, as a compound that promotes dephosphorylation of TAZ and inhibits adipogenesis in C3H10T1/2 cells

J Biochem. 2015 Nov;158(5):413-23. doi: 10.1093/jb/mvv051. Epub 2015 May 14.

Affiliations

¹ Department of Medical Biochemistry, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo 113-8519, Japan;
² Chemical Biology Screening Center.
³ Chemical Biology Screening Center, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo 101-0062, Japan;
⁴ Department of Developmental and Regenerative Biology, Medical Research Institute, Tokyo Medical and Dental University, Tokyo 113-8510, Japan; and.
⁵ Department of Medical Biochemistry, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo 113-8519, Japan; Center for Brain Integration Research, Tokyo Medical and Dental University, Tokyo, 113-8519, Japan yuhammch@tmd.ac.jp.

PMID: 25979969
DOI: 10.1093/jb/mvv051

Abstract

Transcriptional co-activator with PSD-95/Dlg-A/ZO-1 (PDZ)-binding motif (TAZ) regulates in cell proliferation and differentiation. In mesenchymal stem cells it promotes osteogenesis and myogenesis, and suppresses adipogenesis. TAZ activators are expected to prevent osteoporosis, obesity and muscle atrophy. TAZ activation induces epithelial-mesenchymal transition, confers stemness to cancer cells and leads to poor clinical prognosis in cancer patients. In this point of view, TAZ inhibitors should contribute to cancer therapy. Thus, TAZ attracts attention as a two-faced drug target. We screened for TAZ modulators by using human lung cancer A549 cells expressing the fluorescent reporter. Through this assay, we obtained TAZ activator candidates. We unexpectedly found that ethacridine, a widely used antiseptic and abortifacient, enhances the interaction of TAZ and protein phosphatases and increases unphosphorylated and nuclear TAZ. Ethacridine inhibits adipogenesis in mesenchymal C3H10T1/2 cells through the activation of TAZ. This finding suggests that ethacridine is a bona fide TAZ activator and supports that our assay is useful to discover TAZ activators.

Keywords: TAZ; adipogenesis; ethacridine.

Publication types

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

MeSH terms

Active Transport, Cell Nucleus / drug effects
Adaptor Proteins, Signal Transducing / agonists
Adaptor Proteins, Signal Transducing / genetics
Adaptor Proteins, Signal Transducing / metabolism
Adipogenesis / drug effects*
Anti-Obesity Agents / pharmacology*
Cell Line, Tumor
Drug Evaluation, Preclinical
Ethacridine / pharmacology*
Genes, Reporter / drug effects
HEK293 Cells
Humans
Intracellular Signaling Peptides and Proteins / agonists*
Intracellular Signaling Peptides and Proteins / antagonists & inhibitors
Intracellular Signaling Peptides and Proteins / genetics
Intracellular Signaling Peptides and Proteins / metabolism
Luminescent Proteins / genetics
Luminescent Proteins / metabolism
Mesenchymal Stem Cells / cytology
Mesenchymal Stem Cells / drug effects*
Mesenchymal Stem Cells / metabolism
Phosphoproteins / agonists
Phosphoproteins / genetics
Phosphoproteins / metabolism
Phosphorylation / drug effects
Promoter Regions, Genetic / drug effects
Protein Phosphatase 1 / chemistry
Protein Phosphatase 1 / genetics
Protein Phosphatase 1 / metabolism*
Protein Phosphatase 2 / chemistry
Protein Phosphatase 2 / genetics
Protein Phosphatase 2 / metabolism*
Protein Processing, Post-Translational / drug effects
Protein Serine-Threonine Kinases / antagonists & inhibitors
Protein Serine-Threonine Kinases / genetics
Protein Serine-Threonine Kinases / metabolism
RNA Interference
Recombinant Fusion Proteins / chemistry
Recombinant Fusion Proteins / metabolism
Trans-Activators
Transcription Factors
Transcriptional Coactivator with PDZ-Binding Motif Proteins
Tumor Suppressor Proteins / antagonists & inhibitors
Tumor Suppressor Proteins / genetics
Tumor Suppressor Proteins / metabolism
YAP-Signaling Proteins

Substances

Adaptor Proteins, Signal Transducing
Anti-Obesity Agents
Intracellular Signaling Peptides and Proteins
Luminescent Proteins
Phosphoproteins
Recombinant Fusion Proteins
Trans-Activators
Transcription Factors
Transcriptional Coactivator with PDZ-Binding Motif Proteins
Tumor Suppressor Proteins
WWTR1 protein, human
YAP-Signaling Proteins
YAP1 protein, human
LATS1 protein, human
LATS2 protein, human
Protein Serine-Threonine Kinases
PPP1CA protein, human
PPP2CA protein, human
Protein Phosphatase 1
Protein Phosphatase 2
Ethacridine