A highly sensitive reporter system to monitor endogenous YAP1/TAZ activity and its application in various human cells.

Autor: Hikasa H; Department of Biochemistry, School of Medicine, University of Occupational and Environmental Health, Fukuoka, Japan.; Division of Cancer Genetics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan., Kawahara K; Division of Cancer Genetics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.; Department of Molecular Oncology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan., Inui M; Department of Biochemistry, School of Medicine, University of Occupational and Environmental Health, Fukuoka, Japan., Yasuki Y; Department of Molecular Oncology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan.; Department of Chemistry and Bioscience, Graduate School of Science and Engineering, Kagoshima University, Kagoshima, Japan., Yamashita K; Department of Molecular Oncology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan.; Department of Chemistry and Bioscience, Graduate School of Science and Engineering, Kagoshima University, Kagoshima, Japan., Otsubo K; Division of Cancer Genetics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.; Department of Respiratory Medicine, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan., Kitajima S; Department of Biochemistry, School of Medicine, University of Occupational and Environmental Health, Fukuoka, Japan.; Institute for Advanced Biosciences, Keio University, Yamagata, Japan., Nishio M; Division of Cancer Genetics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.; Division of Molecular and Cellular Biology, Kobe University Graduate School of Medicine, Kobe, Japan., Arima K; Department of Chemistry and Bioscience, Graduate School of Science and Engineering, Kagoshima University, Kagoshima, Japan., Endo M; Department of Molecular Biology, University of Occupational and Environmental Health, Fukuoka, Japan., Taira M; Faculty of Science and Engineering, Chuo University, Tokyo, Japan., Suzuki A; Division of Cancer Genetics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.; Division of Molecular and Cellular Biology, Kobe University Graduate School of Medicine, Kobe, Japan.
Jazyk: angličtina
Zdroj: Cancer science [Cancer Sci] 2024 Oct; Vol. 115 (10), pp. 3370-3383. Date of Electronic Publication: 2024 Aug 18.
DOI: 10.1111/cas.16316
Abstrakt: The activation of yes-associated protein 1 (YAP1) and transcriptional co-activator with PDZ-binding motif (TAZ) has been implicated in both regeneration and tumorigenesis, thus representing a double-edged sword in tissue homeostasis. However, how the activity of YAP1/TAZ is regulated or what leads to its dysregulation in these processes remains unknown. To explore the upstream stimuli modulating the cellular activity of YAP1/TAZ, we developed a highly sensitive YAP1/TAZ/TEAD-responsive DNA element (YRE) and incorporated it into a lentivirus-based reporter cell system to allow for sensitive and specific monitoring of the endogenous activity of YAP1/TAZ in terms of luciferase activity in vitro and Venus fluorescence in vivo. Furthermore, by replacing YRE with TCF- and NF-κB-binding DNA elements, we demonstrated the applicability of this reporter system to other pathways such as Wnt/β-catenin/TCF- and IL-1β/NF-κB-mediated signaling, respectively. The practicality of this system was evaluated by performing cell-based reporter screening of a chemical compound library consisting of 364 known inhibitors, using reporter-introduced cells capable of quantifying YAP1/TAZ- and β-catenin-mediated transcription activities, which led to the identification of multiple inhibitors, including previously known as well as novel modulators of these signaling pathways. We further confirmed that novel YAP1/TAZ modulators, such as potassium ionophores, Janus kinase inhibitors, platelet-derived growth factor receptor inhibitors, and genotoxic stress inducers, alter the protein level or phosphorylation of endogenous YAP1/TAZ and the expression of their target genes. Thus, this reporter system provides a powerful tool to monitor endogenous signaling activities of interest (even in living cells) and search for modulators in various cellular contexts.
(© 2024 The Author(s). Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)
Databáze: MEDLINE
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