title: Pituitary adenylate cyclase-activating polypeptide type 1 receptor (PAC1) gene is suppressed by transglutaminase 2 activation.
abstract: Pituitary adenylate cyclase-activating polypeptide (PACAP) functions as a neuroprotective factor through the PACAP type 1 receptor, PAC1. In a previous work, we demonstrated that nerve growth factor augmented PAC1 gene expression through the activation of Sp1 via the Ras/MAPK pathway. We also observed that PAC1 expression in Neuro2a cells was transiently suppressed during in vitro ischemic conditions, oxygen-glucose deprivation (OGD). Because endoplasmic reticulum (ER) stress is induced by ischemia, we attempted to clarify how ER stress affects the expression of PAC1. Tunicamycin, which induces ER stress, significantly suppressed PAC1 gene expression, and salubrinal, a selective inhibitor of the protein kinase RNA-like endoplasmic reticulum kinase signaling pathway of ER stress, blocked the suppression. In luciferase reporter assay, we found that two Sp1 sites were involved in suppression of PAC1 gene expression due to tunicamycin or OGD. Immunocytochemical staining demonstrated that OGD-induced transglutaminase 2 (TG2) expression was suppressed by salubrinal or cystamine, a TG activity inhibitor. Further, the OGD-induced accumulation of cross-linked Sp1 in nuclei was suppressed by cystamine or salubrinal. Together with cystamine, R283, TG2-specific inhibitor, and siRNA specific for TG2 also ameliorated OGD-induced attenuation of PAC1 gene expression. These results suggest that Sp1 cross-linking might be crucial in negative regulation of PAC1 gene expression due to TG2 in OGD-induced ER stress.
authors.affiliation: From the Department of Pharmacology, Graduate School of Medical and Dental Sciences, University of Kagoshima, Kagoshima 890-8544, Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Miura
authors.affiliation: From the Department of Pharmacology, Graduate School of Medical and Dental Sciences, University of Kagoshima, Kagoshima 890-8544, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Kambe
authors.affiliation: From the Department of Pharmacology, Graduate School of Medical and Dental Sciences, University of Kagoshima, Kagoshima 890-8544, Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Inoue
authors.affiliation: the Molecular Ligand Biology Research Team, Chemical Genomics Research Group, Chemical Biology Department, RIKEN Advanced Science Institute, Wako 351-0198, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Tatsukawa
authors.affiliation: From the Department of Pharmacology, Graduate School of Medical and Dental Sciences, University of Kagoshima, Kagoshima 890-8544, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Kurihara
authors.affiliation: the School of Life and Health Sciences, Aston University, Birmingham B4 7ET, United Kingdom.
authors.forename: Martin
authors.initials: M
authors.lastname: Griffin
authors.affiliation: the Molecular Ligand Biology Research Team, Chemical Genomics Research Group, Chemical Biology Department, RIKEN Advanced Science Institute, Wako 351-0198, Japan.
authors.forename: Soichi
authors.initials: S
authors.lastname: Kojima
authors.affiliation: From the Department of Pharmacology, Graduate School of Medical and Dental Sciences, University of Kagoshima, Kagoshima 890-8544, Japan,. Electronic address: amiyata@m3.kufm.kagoshima-u.ac.jp.
authors.forename: Atsuro
authors.initials: A
authors.lastname: Miyata
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D015687:Cell Hypoxia / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D002934:Cinnamates / Q000494:pharmacology; D003538:Cystamine / Q000494:pharmacology; D059865:Endoplasmic Reticulum Stress; D004789:Enzyme Activation / Q000187:drug effects; D004790:Enzyme Induction / Q000187:drug effects / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004794:Enzyme Repression / Q000187:drug effects / Q000235:genetics; D019204:GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D020935:MAP Kinase Signaling System; D051379:Mice; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D051237:Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I / Q000096:biosynthesis* / Q000235:genetics; D020218:Response Elements; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D013890:Thiourea / Q000031:analogs & derivatives / Q000494:pharmacology; D011503:Transglutaminases / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D014415:Tunicamycin / Q000494:pharmacology
chemical_list: C495504:Adcyap1r1 protein, mouse; D000900:Anti-Bacterial Agents; D002934:Cinnamates; D004791:Enzyme Inhibitors; D051237:Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I; D016329:Sp1 Transcription Factor; C496827:salubrinal; D014415:Tunicamycin; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins; D013890:Thiourea; D003538:Cystamine
keywords: ER Stress; G Protein-coupled Receptors (GPCRs); Gene Expression; Ischemia; Neurons; Neuropeptide; PAC1; Pituitary Adenylate Cyclase-activating Polypeptide; Sp1; Transglutaminase 2
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