title: Biphasic Regulation of Yes-associated Protein (YAP) Cellular Localization, Phosphorylation, and Activity by G Protein-coupled Receptor Agonists in Intestinal Epithelial Cells: A NOVEL ROLE FOR PROTEIN KINASE D (PKD).
abstract: We examined the regulation of Yes-associated protein (YAP) localization, phosphorylation, and transcriptional activity in intestinal epithelial cells. Our results show that stimulation of intestinal epithelial IEC-18 cells with the G protein-coupled receptor (GPCR) agonist angiotensin II, a potent mitogen for these cells, induced rapid translocation of YAP from the nucleus to the cytoplasm (within 15 min) and a concomitant increase in YAP phosphorylation at Ser(127) and Ser(397) Angiotensin II elicited YAP phosphorylation and cytoplasmic accumulation in a dose-dependent manner (ED50 = 0.3 nm). Similar YAP responses were provoked by stimulation with vasopressin or serum. Treatment of the cells with the protein kinase D (PKD) family inhibitors CRT0066101 and kb NB 142-70 prevented the increase in YAP phosphorylation on Ser(127) and Ser(397) via Lats2, YAP cytoplasmic accumulation, and increase in the mRNA levels of YAP/TEAD-regulated genes (Ctgf and Areg). Furthermore, siRNA-mediated knockdown of PKD1, PKD2, and PKD3 markedly attenuated YAP nuclear-cytoplasmic shuttling, phosphorylation at Ser(127), and induction of Ctgf and Areg expression in response to GPCR activation. These results identify a novel role for the PKD family in the control of biphasic localization, phosphorylation, and transcriptional activity of YAP in intestinal epithelial cells. In turn, YAP and TAZ are necessary for the stimulation of the proliferative response of intestinal epithelial cells to GPCR agonists that act via PKD. The discovery of interaction between YAP and PKD pathways identifies a novel cross-talk in signal transduction and demonstrates, for the first time, that the PKDs feed into the YAP pathway.
authors.affiliation: From the Division of Digestive Diseases, Department of Medicine, David Geffen School of Medicine.
authors.forename: Jia
authors.initials: J
authors.lastname: Wang
authors.affiliation: From the Division of Digestive Diseases, Department of Medicine, David Geffen School of Medicine, CURE: Digestive Diseases Research Center, and the Veterans Affairs Greater Los Angeles Health Care System, Los Angeles, California 90073.
authors.forename: James
authors.initials: J
authors.lastname: Sinnett-Smith
authors.affiliation: From the Division of Digestive Diseases, Department of Medicine, David Geffen School of Medicine.
authors.forename: Jan V
authors.initials: JV
authors.lastname: Stevens
authors.affiliation: From the Division of Digestive Diseases, Department of Medicine, David Geffen School of Medicine, the Veterans Affairs Greater Los Angeles Health Care System, Los Angeles, California 90073.
authors.forename: Steven H
authors.initials: SH
authors.lastname: Young
authors.affiliation: From the Division of Digestive Diseases, Department of Medicine, David Geffen School of Medicine, CURE: Digestive Diseases Research Center, and the Veterans Affairs Greater Los Angeles Health Care System, Los Angeles, California 90073 Molecular Biology Institute, UCLA, Los Angeles, California 90095 and erozengurt@mednet.ucla.edu.
authors.forename: Enrique
authors.initials: E
authors.lastname: Rozengurt
mesh_terms: D000217:Acyltransferases; D016564:Amyloid beta-Protein Precursor / Q000235:genetics / Q000378:metabolism; D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism*; D003593:Cytoplasm / Q000235:genetics / Q000378:metabolism*; D020895:Enterocytes / Q000378:metabolism*; D006575:Heterocyclic Compounds, 3-Ring / Q000494:pharmacology; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects / Q000235:genetics; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport / Q000187:drug effects / Q000235:genetics; D011743:Pyrimidines / Q000494:pharmacology; D051381:Rats; D043562:Receptors, G-Protein-Coupled / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D013841:Thiazepines / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D000091102:YAP-Signaling Proteins
chemical_list: D016564:Amyloid beta-Protein Precursor; D051017:Apoptosis Regulatory Proteins; C551536:CRT 0066101; C437330:CTFgamma protein, rat; D006575:Heterocyclic Compounds, 3-Ring; D010446:Peptide Fragments; D011743:Pyrimidines; D043562:Receptors, G-Protein-Coupled; D013841:Thiazepines; D014157:Transcription Factors; D000091102:YAP-Signaling Proteins; C509425:Yap1 protein, rat; C579279:kb-NB142-70; D000217:Acyltransferases; C544439:Tafazzin protein, rat; C089212:protein kinase D; C000591485:Lats2 protein, rat; D017346:Protein Serine-Threonine Kinases; D011493:Protein Kinase C
keywords: G protein-coupled receptor (GPCR); angiotensin II; protein kinase C (PKC); protein kinase D (PKD); yes-associated protein (YAP)