Pituitary adenylate cyclase-activating polypeptide type 1 receptor (PAC1) gene is suppressed by transglutaminase 2 activation

Ayako Miura, Yuki Kambe, Kazuhiko Inoue, Hideki Tatsukawa, Takashi Kurihara, Martin Griffin, Soichi Kojima, Atsuro Miyata

Research output: Contribution to journalArticle

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. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

Original languageEnglish
Pages (from-to)32720-32730
Number of pages11
JournalJournal of Biological Chemistry
Volume288
Issue number45
Early online date17 Sep 2013
DOIs
Publication statusPublished - 8 Nov 2013

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Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I
Endoplasmic Reticulum Stress
Gene expression
Genes
Chemical activation
Cystamine
Oxygen
Glucose
Gene Expression
Tunicamycin
Pituitary Adenylate Cyclase-Activating Polypeptide
Gene Expression Regulation
Nerve Growth Factor
Protein Kinase Inhibitors
Luciferases
Endoplasmic Reticulum
Small Interfering RNA
transglutaminase 2
Assays
Phosphotransferases

Bibliographical note

© 2013 by The American Society for Biochemistry and Molecular Biology, Inc

Keywords

  • ER stress
  • G protein coupled receptors
  • gene expression
  • ischemia
  • neurons
  • transglutaminase 2
  • pituitary adenylate cyclase-activating polypeptide
  • PAC1
  • Sp1
  • neuropeptide

Cite this

Miura, Ayako ; Kambe, Yuki ; Inoue, Kazuhiko ; Tatsukawa, Hideki ; Kurihara, Takashi ; Griffin, Martin ; Kojima, Soichi ; Miyata, Atsuro. / Pituitary adenylate cyclase-activating polypeptide type 1 receptor (PAC1) gene is suppressed by transglutaminase 2 activation. In: Journal of Biological Chemistry. 2013 ; Vol. 288, No. 45. pp. 32720-32730.
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Pituitary adenylate cyclase-activating polypeptide type 1 receptor (PAC1) gene is suppressed by transglutaminase 2 activation. / Miura, Ayako; Kambe, Yuki; Inoue, Kazuhiko; Tatsukawa, Hideki; Kurihara, Takashi; Griffin, Martin; Kojima, Soichi; Miyata, Atsuro.

In: Journal of Biological Chemistry, Vol. 288, No. 45, 08.11.2013, p. 32720-32730.

Research output: Contribution to journalArticle

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