title: A cell-based assay to screen stimulators of the Hippo pathway reveals the inhibitory effect of dobutamine on the YAP-dependent gene transcription.
abstract: The mammalian Hippo pathway is composed of mammalian Ste20-like (MST) kinases and large tumour suppressor (LATS) kinases. Upon the activation of the pathway, MST kinases phosphorylate and activate LATS kinases, which in turn phosphorylate transcriptional co-activators, yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ), recruit them to the cytosol from the nucleus and turn off cell cycle-promoting and anti-apoptotic gene transcriptions. Thus, the pathway restricts cell overgrowth and prevents tumourigenesis. Although a high cell density and stress signallings are known to activate the pathway, no specific stimulators are so far reported. As the dysfunction of the pathway is frequent in human cancers and correlates with poor prognosis, it is important to find out reagents that stimulate the pathway for not only basic research but also clinical medicine. We here developed a cell-based method of screening reagents that induce the recruitment of YAP to the cytosol. Using this method, we found that dobutamine inhibits the YAP-dependent gene transcription. Contrary to our expectations, the effect of dobutamine is independent of the Hippo pathway but our method opens the possibility to discover Hippo pathway stimulators or Hippo-independent YAP inhibitors.
authors.affiliation: Department of Medical Biochemistry, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo 113-8519, Japan.
authors.forename: Yijun
authors.initials: Y
authors.lastname: Bao
authors.forename: Kentaro
authors.initials: K
authors.lastname: Nakagawa
authors.forename: Zeyu
authors.initials: Z
authors.lastname: Yang
authors.forename: Mitsunobu
authors.initials: M
authors.lastname: Ikeda
authors.forename: Kanchanamala
authors.initials: K
authors.lastname: Withanage
authors.forename: Mari
authors.initials: M
authors.lastname: Ishigami-Yuasa
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Okuno
authors.forename: Shoji
authors.initials: S
authors.lastname: Hata
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nishina
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Hata
mesh_terms: D000217:Acyltransferases; D018797:Cell Cycle Proteins; D045744:Cell Line, Tumor; D004280:Dobutamine / Q000494:pharmacology*; D004354:Drug Screening Assays, Antitumor; D057809:HEK293 Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D034741:RNA, Small Interfering; D011941:Receptors, Adrenergic / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014157:Transcription Factors / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000099317:Hippo Kinases
chemical_list: D000217:Acyltransferases; D018797:Cell Cycle Proteins; D004280:Dobutamine; D006861:Hydrogen Peroxide; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D017346:Protein Serine-Threonine Kinases; D034741:RNA, Small Interfering; D011941:Receptors, Adrenergic; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C456659:YY1AP1 protein, human; C108675:TAFAZZIN protein, human; C092745:LATS1 protein, human; C109163:STK24 protein, human; D000099317:Hippo Kinases; C406449:LATS2 protein, human